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Tuberculous lymphadenitis (TBL) is defined by reduced proinflammatory cytokines and elevated CD4 + , CD8 + T cells and decreased CD8 + cytotoxic markers. However, ex-vivo phenotyping of diverse leucocytes in TBL has not been done. We show activated and atypical B cells, myeloid dendritic cells (mDCs), classical, non-classical and intermediate monocytes, T regulatory (T regs) cells, CD4 + T cell effector memory RA (TEMRA), CD4 + effector and CD8 + central memory phenotypes were significantly increased in TBL compared to LTB individuals. In contrast, classical memory and plasma B cells, plasmacytoid DCs (pDCs), CD8 + TEMRA, CD4 + naïve and central memory cells were significantly decreased in TBL compared to LTB individuals. Some of the leucocyte frequencies (atypical memory B cells, pDCs, myeloid-derived suppressor cells, CD4 + effector and CD8 + central memory was increased; activated memory and plasma B cell, mDCs, classical, non-classical, intermediate monocytes, T regs, CD4 + TEMRA, CD4 + , CD8 + naïve and effector memory cells and CD8 + central memory cells were decreased) were significantly modulated after anti-TB treatment among TBL individuals. UMAP analysis show that leucocyte subsets or islands expressing specific markers were significantly different in TBL baseline and post-treatment individuals. Overall, we suggest altered frequencies of diverse leucocytes influences the disease pathology and protective immunity in TBL individuals., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Granuloma formation is the pathologic hallmark of tuberculosis (TB). Few studies have detailed the exact production of cytokines in human granulomatous inflammation and little is known about accessory molecule expressions in tuberculous granulomas. We aimed to identify some of the components of the immune response in granulomas in HIV-positive and -negative lymph nodes. We investigated the immunohistochemical profiles of CD4+, CD8+, CD68+, Th-17, Forkhead box P3 (FOXP3) cells, accessory molecule expression (human leukocyte antigen [HLA] classes I and II), and selected cytokines (interleukins 2, 4, and 6 and interferon-γ) of various cells, in granulomas within lymph nodes from 10 HIV-negative (-) and 10 HIV-positive (+) cases. CD4+ lymphocyte numbers were retained in HIV- granulomas, whereas CD4+:CD8 + cell were reversed in HIV+ TB granulomas. CD68 stained all histiocytes. Granulomas from the HIV+ group demonstrated a significant increase in FOXP3 cells. Interleukin-2 cytoplasmic expression was similar in both groups. Interferon-gamma (IFN-γ) expression was moderately increased, IL-6 was statistically increased and IL-4 expression was marginally lower in cells from HIV- than HIV+ TB granulomas. Greater numbers of cells expressed IFN-γ and IL-6 than IL-2 and IL-4 in HIV- TB granulomas. This study highlights the varied cytokine production in HIV-positive and -negative TB granulomas and indicates the need to identify localized tissue factors that play a role in mounting an adequate immune response required to halt infection. Although TB mono-infection causes variation in cell marker expression and cytokines in granulomas, alterations in TB and HIV coinfection are greater, pointing toward evolution of microorganism synergism.

Background: Tuberculosis (TB) though primarily affects the lungs it may also affect the other parts of the body and referred as extra pulmonary (EPTB). This study is focused on understanding the genetic diversity and molecular epidemiology of Mycobacterium tuberculosis (M.tb) among tuberculous lymphadenitis (TBL), a form of EPTB patients identified in Chennai, Tamil Nadu., Methods: The genetic diversity was identified by performing spoligotyping on the M.tb clinical isolates that were recovered from lymph node samples. A total of 71 M.tb isolates were recovered from extra pulmonary lymph node samples and subjected to Drug susceptibility testing and spoligotyping was carried out. In addition, immunological characterization from blood of same individuals from whom M.tb was isolated was carried out between the two major lineages groups East African Indian 3 (EAI3) and non-EAI3 strains by ELISA. The results of spoligotyping patterns were compared with the world Spoligotyping Database of Institute Pasteur de Guadeloupe (SpolDB4)., Results: We found 41 spoligotype patterns and their associated lineages. Out of 41 spoligotype pattern, only 22 patterns are available in the spoldB4 database with Spoligotype international Type (SIT) number and remaining patterns were orphan strains without SIT number. The most predominant spoligotype lineage that was found in lymph node sample in this region of India was EAI (36), followed by central Asian strain (CAS) (6), T1 (5), Beijing (3), Latin American & Mediterranean (LAM) (2), U (1), X2 (1) and orphan (22). In addition to EAI, CAS and Beijing, our study identified the presence of orphan and unique spoligotyping patterns in Chennai region. We observed six drug resistant isolates. Out of six drug resistant isolates, four were resistant to isoniazid drug and associated with EAI family. Moreover, we observed increased levels of type 2 and type 17 cytokine profiles between EAI3 and non-EAI family, infected individuals., Conclusions: The study confirms that EAI lineage to be the most predominant lineages in EPTB patients with lymphadenitis and were found to have increased type 1 and type 17 proinflammatory cytokine profiles.

Tuberculous lymphadenitis (TBL) individuals exhibit reduced frequencies of CD8 + T cells expressing cytotoxic markers in peripheral blood. However, the frequencies of cytotoxic marker expressing CD4 + , CD8 + T cells, and NK cells at the site of infection is not known. Therefore, we measured the baseline and mycobacterial antigen specific frequencies of cytotoxic markers expressing CD4 + , CD8 + T cells, and NK cells in the LN ( n = 18) and whole blood ( n = 10) of TBL individuals. TBL LN is associated with lower frequencies of CD4 + T cells expressing cytotoxic markers (Granzyme B, CD107a) compared to peripheral blood at baseline and in response to PPD, ESAT-6, and CFP-10 antigen stimulation. Similarly, lower frequencies of CD8 + T cells expressing cytotoxic markers (Perforin, Granzyme B, and CD107a) were also present in the TBL LN at baseline and following (except perforin) antigen stimulation. Finally, at baseline and after antigen (PPD, ESAT-6, and CFP-10) stimulation, frequencies of NK cells expressing cytotoxic markers were also significantly lower in TBL LN compared to whole blood. Hence, TBL is characterized by diminished frequencies of cytotoxic marker expressing CD4 + , CD8 + T cells, and NK cells at the site of infection, which might reflect the lack of protective immune responses at the site of Mycobacterium tuberculosis infection., (Copyright © 2020 Kathamuthu, Moideen, Sridhar, Baskaran and Babu.)

Tuberculosis (TB) is a serious public health problem in Bangladesh. National tuberculosis control program recognizes that almost half of the TB cases remain undiagnosed in the country. To increase case detection rate, it is very important to familiarize the physicians with unusual presentation of TB. We describe a 51 years old woman with a past medical history of Hypertension (HTN), Type 2 Diabetes Mellitus (DM), and Nonalcoholic steatohepatitis-chronic liver disease (NASH-CLD) who presented to us with low grade fever, anorexia, nausea, and recurrent vomiting for one month. Physical examination and laboratory tests revealed no significant abnormalities and symptoms were treated symptomatically. After about two months, the condition did not improve. All routine blood biochemistry and imaging reports were not suggestive of any disease except for high ESR and abnormal LFT (mild increase in ALP, ALT and moderate increase in GGTP). To exclude the differential diagnoses (such as abdominal TB), we advised computed tomography (CT) scan of chest and abdomen but the results came out normal. Her PPD came out positive but it was not confirmatory of TB as the patient was previously vaccinated with BCG vaccine. As the patient was immune-compromised we suggested starting Anti-TB drugs based on clinical judgment and in the context of Bangladesh being a TB endemic area. But the patient was not convinced to take anti-Tb drugs without definite diagnosis. After another month of persistent symptoms a repeat CT of the chest was advised that revealed multiple enlarged mediastinal lymph-nodes. As the patient had a history of CLD and high PT, Fine Needle Aspiration Cytology (FNAC) was deferred. Patient was started on Anti tubercular treatment and symptoms subsided within three weeks. Treatment was continued for one year. This case summarizes the unusual presentation of mediastinal lymph node Tuberculosis in an adult., Competing Interests: Declaration of Competing Interest All authors have none to declare., (Copyright © 2020 Tuberculosis Association of India. Published by Elsevier B.V. All rights reserved.)

Filarial infections are known to modulate cytokine responses in pulmonary tuberculosis by their propensity to induce Type 2 and regulatory cytokines. However, very little is known about the effect of filarial infections on extra-pulmonary forms of tuberculosis. Thus, we have examined the effect of filarial infections on the plasma levels of various families of (IL-1, IL-12, γC, and regulatory) cytokines and (CC and CXC) chemokines in tuberculous lymphadenitis coinfection. We also measured lymph node culture grades in order to assess the burden of Mycobacterium tuberculosis in the two study groups [Fil+ ( n = 67) and Fil- ( n = 109)]. Our data reveal that bacterial burden was significantly higher in Fil+ compared to Fil- individuals. Plasma levels of IL-1 family (IL-1α, IL-β, IL-18) cytokines were significantly lower with the exception of IL-33 in Fil+ compared to Fil- individuals. Similarly, plasma levels of IL-12 family cytokines -IL-12 and IL-23 were significantly reduced, while IL-35 was significantly elevated in Fil+ compared to Fil- individuals. Filarial infection was also associated with diminished levels of IL-2, IL-9 and enhanced levels of IL-4, IL-10, and IL-1Ra. Similarly, the Fil+ individuals were linked to elevated levels of different CC (CCL-1, CCL-2, CCL-3, CCL-11) and CXC (CXCL-2, CXCL-8, CXCL-9, CXCL-11) chemokines. Therefore, we conclude that filarial infections exert powerful bystander effects on tuberculous lymphadenitis, effects including modulation of protective cytokines and chemokines with a direct impact on bacterial burdens., (Copyright © 2020 Kathamuthu, Munisankar, Banurekha, Nair, Sridhar and Babu.)

Tuberculous lymphadenitis (TBL) is associated with the expansion of CD4 + and CD8 + T cells expressing Type 1 and Type 17 cytokines in the peripheral blood. However, the expression pattern of cytokine producing natural killer (NK) cells in both the peripheral blood and affected lymph nodes i.e. site of infection in TBL have not been examined. Hence, we have analyzed the baseline and mycobacterial antigen specific NK cell cytokine frequencies in whole blood of TBL and pulmonary tuberculosis (PTB) individuals. We have also examined the NK cell frequencies before and after treatment completion and in peripheral blood versus affected lymph nodes (LN) of TBL individuals. TBL is characterized by diminished frequencies of NK cells expressing Type 1 (IFNγ, TNFα), Type 17 (IL-17F) cytokines compared to PTB individuals upon antigen-specific stimulation. In contrast, TBL individuals did not exhibit any significant differences in the frequencies of NK cells expressing Type 1 and Type 17 cytokines upon completion of anti-tuberculosis treatment. LN of TBL is associated with altered frequencies of NK cells expressing Type 17 (increased IL-17F and decreased IL-22) cytokines when compared to peripheral blood. Thus, we conclude that TBL individuals are characterized by diminished frequencies of NK cells expressing Type 1/Type 17 cytokines., (Copyright © 2019. Published by Elsevier Ltd.)

Lymph node tuberculosis (LNTB) is characterized by the enhanced baseline and antigen-specific production of type 1/17 cytokines and reduced baseline and antigen-specific production of interleukin (IL)-1β and IL-18 at the site of infection when compared with peripheral blood. However, the cytokine profile in the lymph nodes (LNs) of Mycobacterium tuberculosis culture-positive LNTB (LNTB+) and negative LNTB (LNTB-) has not been examined. To address this, we have examined the baseline and mycobacterial antigen-stimulated cytokine levels of type 1 (interferon gamma [IFNγ], tumor necrosis factor alpha [TNFα], IL-2), type 2 (IL-4, IL-5, and IL-13), type 17 (IL-17A, IL-17F, and IL-22), pro-inflammatory (IL-1α, IL-1β, IL-18, and granulocyte macrophage colony-stimulating factor [GM-CSF]), and regulatory cytokines (IL-10, transforming growth factor beta [TGF-β]) cytokines in the LN culture supernatants of LNTB+ and LNTB- individuals. We have observed significantly enhanced baseline levels of IL-13 and IL-10 and significantly reduced baseline levels of IL-4 and GM-CSF in LNTB+ individuals compared with LNTB- individuals. By contrast, we have observed significantly enhanced levels of type 1 (IFNγ, TNFα, and IL-2), type 17 (IL-17F and IL-22), and pro-inflammatory (IL-1α and GM-CSF) cytokines and significantly reduced levels of TGFβ in response to purified protein derivative, early secreted antigen-6, and culture filtrate protein-10 antigens in LNTB+ compared with LNTB- individuals. On phorbol 12-myristate 13-acetate/ionomycin stimulation, no significant difference was observed for any of the cytokines examined. Thus, our study revealed several interesting differences in the cytokine profiles of mycobacterial antigen-stimulated LN cultures in LNTB+ and LNTB- individuals. Therefore, we suggest the presence of mycobacteria plays a significant role in driving the cytokine response at the site of infection in LNTB.

Background: Helminth infections are known to regulate cytokine responses in both pulmonary and latent tuberculosis infection. Whether helminth infections also modulate cytokine responses in extra-pulmonary tuberculosis, specifically tuberculous lymphadenitis (TBL), has not been examined thus far., Methodology: Hence, to determine the cytokine profile in helminth-TBL coinfection, we measured the systemic and mycobacterial (TB)-antigen stimulated levels of Type 1, Type 2, Type 17, regulatory and pro-inflammatory cytokines in TBL individuals coinfected with or without Strongyloides stercoralis (Ss) infection., Significant Findings: TBL-Ss+ individuals have significantly higher bacterial burdens in the affected lymph nodes in comparison to TBL-Ss- individuals. TBL-Ss+ individuals exhibit significantly enhanced plasma levels of Type 2 (IL-5 and IL-13), Type 17 (IL-17 and IL-22) and regulatory (IL-10) cytokines in comparison to TBL-Ss- individuals. In contrast, TBL-Ss+ individuals exhibit significantly diminished plasma levels of pro-inflammatory cytokines (IL-1α and GM-CSF) in comparison to TBL-Ss- individuals. TBL-Ss+ individuals also exhibit significantly diminished unstimulated or mycobacterial-antigen stimulated levels of Type 1, Type 17 or IL-1 family cytokines in comparison to TBL-Ss- individuals but no differences in mitogen stimulated cytokine levels., Conclusion: Therefore, our data reveal a profound influence of Ss infection on the bacteriological profile of TBL and suggesting that the underlying modulation of cytokine responses might be a mechanism by which this helminth infection could impart a detrimental effect on the pathogenesis of TBL disease., Competing Interests: The authors have declared that no competing interests exist.

Background: Lymph node tuberculosis (LNTB) is the most frequent extrapulmonary form of tuberculosis (TB). Studies of human tuberculosis at sites of disease are limited. LNTB provides a unique opportunity to compare local in situ and peripheral blood immune response in active Mycobacterium tuberculosis (Mtb) disease. The present study analysed T regulatory cells (Treg) frequency and activation along with CD4+ T cell function in lymph nodes from LNTB patients., Results: Lymph node mononuclear cells (LNMC) were compared to autologous peripheral blood mononuclear cells (PBMC). LNMC were enriched for CD4+ T cells with a late differentiated effector memory phenotype. No differences were noted in the frequency and mutifunctional profile of memory CD4+ T cells specific for Mtb. The proportion of activated CD4+ and Tregs in LNMC was increased compared to PBMC. The correlation between Tregs and activated CD4+ T cells was stronger in LNMC than PBMC. Tregs in LNMC showed a strong positive correlation with Th1 cytokine production (IL2, IFNγ and TNFα) as well as MIP-1α after Mtb antigen stimulation. A subset of Tregs in LNMC co-expressed HLA-DR and CD38, markers of activation., Conclusion: Further research will determine the functional relationship between Treg and activated CD4+ T cells at lymph node sites of Mtb infection.

Rational: Granulomatous polyangiitis (GPA) is a type of vasculitis involving medium and small arteries, typically affecting the upper and lower respiratory tract with coexisting glomerulonephritis. GPA is also characterized by necrotizing granulomatous inflammation and the presence of antineutrophil cytoplasm antibodies (ANCA). So far, various infections have lead to elevation of titers of serum ANCA, making it difficult to diagnose., Patient Concerns: We report a 50-year-old woman who was diagnosed as tuberculous lymphadenitis. During the treatment by anti-tuberculosis (TB) drugs, rapidly progressive renal failure and pleurisy had appeared with elevated titer of PR3-ANCA. Renal biopsy revealed crescentic glomerulonephritis., Diagnosis: Renal biopsy revealed crescentic glomerulonephritis and diagnosis of GPA was made., Interventions: Steroid therapy had been started with continuation of anti-TB drugs., Outcomes: Renal dysfunction had gradually recovered and pleurisy had disappeared with decreasing titer of PR3-ANCA., Lessons: This is the first report of GPA complicated by TB infection. When we encounter a case with rapidly progressive renal failure during the TB infection, complication of GPA should be suspected as 1 of the different diagnosis.

Tuberculous lymphadenitis (TBL) is characterized by an expansion of Th1 and Th17 cells with altered serum levels of proinflammatory cytokines. However, the cytokine profile at the site of infection, i.e., the affected lymph nodes, has not been examined in detail. To estimate the baseline and mycobacterial antigen-stimulated concentrations of type 1, type 17, and other proinflammatory cytokines in patients with TBL ( n = 14), we examined both the baseline and the antigen-specific concentrations of these cytokines before and after chemotherapy and compared them with those in individuals with pulmonary tuberculosis (PTB) ( n = 14). In addition, we also compared the cytokine responses in whole blood and those in the lymph nodes of TBL individuals. We observed significantly enhanced baseline and antigen-specific levels of type 1 cytokines (gamma interferon [IFN-γ] and tumor necrosis factor alpha [TNF-α]) and a type 17 cytokine (interleukin-17 [IL-17]) and significantly diminished baseline and antigen-specific levels of proinflammatory cytokines (IL-1β and IL-18) in the whole blood of TBL individuals compared to those in the whole blood of PTB individuals. Moreover, we also observed a pattern of baseline and antigen-specific cytokine production at the site of infection (lymph node) similar to that in the whole blood of TBL individuals. Following standard antituberculosis (anti-TB) treatment, we observed alterations in the baseline and/or antigen-specific levels of IFN-γ, TNF-α, IL-1β, and IL-18. TBL is therefore characterized by enhanced baseline and antigen-specific production of type 1 and type 17 cytokines and reduced baseline and antigen-specific production of IL-1β and IL-18 at the site of infection., (Copyright © 2017 American Society for Microbiology.)

Tuberculosis is one of the leading infections after renal transplant, particularly in developing countries where the incidence and prevalence in the general population are high. Diagnosis requires bacteriologic and histologic confirmation. Interactions among the antitubercular drugs and the immunosuppressive agents have to be considered while prescribing, and surveillance for adverse effects is required. Although rare, case reports are available on extrapulmonary tuberculosis in allograft recipients. Here, we present a 25-year-old kidney transplant recipient who was diagnosed with lymph node tuberculosis under uncommon circumstances but who had a good outcome. This case report illustrates the difficulties in diagnosis of tuberculosis, changes in therapeutic protocols, and prognostic factors and highlights the effects of infectious complications with immunosuppressive therapy in this particular patient population.

Background: Type 1, Type 17 and other pro-inflammatory cytokines are known to play an important role in resistance to pulmonary tuberculosis. The role of these cytokines in tuberculous lymphadenitis (TBL) is not well characterized., Methods: To estimate the systemic and mycobacterial antigen - stimulated cytokine concentrations of Type 1, Type 17, other pro-inflammatory and regulatory cytokines in TBL, we examined both the systemic and the antigen-specific concentrations of these cytokines in TBL (n=31) before and after chemotherapy, and compared them with those with latent tuberculosis infection (LTB, n=31)., Results: We observed significantly reduced systemic concentrations of the pro-inflammatory cytokines - IL-1β and IL-18 but not other Type 1 or Type 17 cytokines in TBL compared to LTB. Following standard anti-tuberculosis (TB) treatment, we observed a significant increase in the concentrations of both IL-1β and IL-18. In addition, we also observed significantly reduced baseline or mycobacterial - antigen or mitogen stimulated concentrations of IL-1β and IL-18 in TBL individuals. Similar to systemic cytokine concentrations, anti-TB treatment resulted in significantly increased concentrations of these cytokines following antigen stimulation., Conclusions: TBL is therefore, characterized by reduced systemic and antigen-specific concentrations of IL-1β and IL-18, which are reversible following anti-TB treatment, indicating that these cytokines are potential correlates of protective immunity in TBL., (Published by Elsevier Ltd.)

Objective: The aim of this work was to study the microscopic patterns of human immunodeficiency virus (HIV) lymphadenitis on fine-needle aspiration cytology (FNAC) and correlate them with cluster of differentiation 4 (CD4) counts., Study Design: A retrospective study of known HIV-positive patients who underwent lymph node FNAC over a period of 5 years (2009-2013) was undertaken. The cytology slides were retrieved and reviewed. Out of 317 cases, 38 (11.7%) were diagnosed as HIV lymphadenitis. We analysed the cytomorphological patterns of HIV lymphadenitis and correlated them with the CD4 cell counts., Results: Smears of HIV lymphadenitis were classified akin to histology patterns (A, B, and C) depending on cellularity, number of tingible body macrophages, mitosis, apoptotic bodies, plasma cells, Warthin-Finkeldey giant cells, and proliferating blood vessels. Thirty-one cases showed pattern A, 3 showed pattern B, and 4 were of pattern C. Pattern A had the highest CD4 cell count., Conclusion: Histologic patterns of HIV lymphadenitis are recognisable on FNAC smears. These can offer a clue to the diagnosis and guide further workup, even in the absence of history. The changes can mimic those of the infective lymphadenitis, Castleman disease, and lymphoma. Hence, the clinical history, serological correlation, and awareness of cytomorphology can aid the correct diagnosis., (© 2016 S. Karger AG, Basel.)

Objectives: To study the cytomorphological spectrum of lymph node lesions in HIV-positive patients and correlate the cytological findings with the CD4 count., Study Design: This was a cross-sectional study of 23 months' duration which included 110 HIV-positive cases proved according to the guidelines of the National AIDS Control Organisation. Fine-needle aspiration cytology (FNAC) was done on clinically palpable lymph nodes. Special stains and culture were done wherever necessary. CD4 count was done by flow cytometry and subsequently correlated with the cytological findings., Results: Our study included 110 cases, ranging in age from 6 to 70 years, peaking in the 3rd to 4th decade (n = 46). The male-to-female ratio was 1.75:1 and the predominant site was the cervical group of lymph nodes (n = 71). Each lesion was correlated with CD4 count, laboratory and clinical findings, and was further segregated based on WHO and CDC staging. Cytological lesions were tubercular (53.6%), reactive (27.1%), suppurative (6.4%) lymphadenitis, lymphoma (4.5%), cystic lymphoid hyperplasia (2.8%), metastases (1.9%), cryptococcal lymphadenitis (0.9%). The mean CD4 count was 217.4, 434.4, 181.4, 149, 580, 225, and 207 cells/µL, respectively. There was a highly significant correlation of cytological findings with CD4 count (χ2 value = 44.57 and p < 0.001)., Conclusion: FNAC is a primary, safe, and valuable tool for the identification of opportunistic infections, neoplastic and nonneoplastic lesions, as well as therapeutic modality in certain conditions. Correlation of lesions with CD4 count provides information about immune status, HIV stage and segregating cases, and also aids further evaluation and management., (© 2016 S. Karger AG, Basel.)

The aim of the study was to evaluate the performance of interferon-γ (IFN-γ) release assay (IGRA) (T-SPOT.TB) for patients with suspected lymph node tuberculosis (TB). Of the 405 patients with suspected lymph node TB, enrolled from Beijing Chest Hospital between July 2011 and April 2015, 83 (20.5%) were microbiologically/histopathologically confirmed lymph node TB, and 282 (69.6%) did not have active TB. The remaining 21 inconclusive TB and 19 clinical TB were excluded from the final analysis (9.9%). T-SPOT.TB using peripheral blood mononuclear cells was performed to examine the IFN-γ response to the Mycobacterium tuberculosis-specific antigens early secretory antigenic target 6 and culture filtrate protein 10. The overall sensitivity and specificity for T-SPOT.TB were 90.4% and 70.5%, respectively. Spot-forming cells in the lymph node TB group (184 [48-596/10(6) peripheral blood mononuclear cells {PBMCs}]) were significantly higher than that in the nonactive TB group (0 [0-41]/10(6) PBMCs) (P<0.001). These results suggest that the IGRA assay could be a useful aid in the diagnosis of lymph node TB., (Copyright © 2016. Published by Elsevier Inc.)

Background: Paradoxical tuberculosis (TB)-associated Immune Reconstitution Inflammatory Syndrome (IRIS) is a common complication of combination antiretroviral treatment (cART) initiation in adults residing in resource-limited regions. Little is known about the burden and presentation of TB-IRIS in children initiating cART while receiving TB treatment., Methods: Prospective cohort study of South African children initiating cART while on TB treatment. Children were assessed clinically and by chest x-ray before starting cART and at 2, 4, 6, and 12 weeks post cART initiation. All children who presented with any signs or symptoms suggestive of paradoxical TB-IRIS were classified according to the consensus adult TB-IRIS case definition developed by the International Network for Study of HIV-associated IRIS (INSHI) and reviewed by an independent expert panel., Results: In 7 of the 104 children enrolled in the cohort, symptoms and/or clinical or radiological signs suggestive of paradoxical TB-IRIS developed after a median of 14 days of cART. In two of these cases, there was agreement between the INSHI case definition and the expert panel. In an additional 3 cases, the INSHI criteria were fulfilled but the expert panel made an alternative diagnosis of pneumonia (n = 2) and poor adherence to cART (n = 1)., Conclusions: The burden of paradoxical TB-IRIS in children with underlying TB initiating cART is low. Including response to antibiotic treatment for pneumonia as a criterion for an alternative diagnosis may improve the specificity of the INSHI case definition., (© 2015 Wiley Periodicals, Inc.)

Objectives: The present study was performed to evaluate various cytological patterns and acid fast bacillus (AFB) grades in HIV-infected patients with tuberculous lymphadenitis and to correlate these with each other as well as with peripheral CD4+ T-cell counts., Study Design: Ninety-two HIV-seropositive patients, cytologically diagnosed with tuberculous lymphadenitis, were evaluated. Fine needle aspiration cytology was performed as an outpatient procedure. Sonographic guidance was sought for internally sited lymph nodes. Cytopathological details were assessed on routinely stained and Ziehl-Neelsen-stained smears. Appropriate AFB grades were assigned. CD4+ T-cell counts were obtained immediately. Finally, the cytopathological findings, AFB grades and CD4+ T-cell counts were corroborated with each other., Results: Epithelioid cell granuloma in the presence of caseation appeared to be the most frequent (66.3%) cytomorphology on aspirated smears. AFB grades 3+ (37%) and 4+ (35.9%) were the commonest patterns of bacillary involvement. The mycobacterial density and cytological features significantly correlated with CD4+ T-cell counts., Conclusions: In HIV-associated tuberculous lymphadenitis, AFB grade and CD4+ T-cell counts worsen with the appearance of necrosis. Here, the peripheral CD4+ T-cell counts inversely correlated with bacillary load. Collectively, peripheral CD4+ T-cell counts, cytological findings and AFB grade exemplify the immune status in these patients., (© 2015 S. Karger AG, Basel.)

Abdominal tuberculosis manifesting as isolated lymphadenopathy is rare, particularly in children. Tuberculous involvement of the pancreatic head and peripancreatic area can simulate a neoplasm of the pancreatic head. To our knowledge, obstructive jaundice caused by tuberculous lymphadenopathy has not been reported in children or adolescents. Here we present radiologic findings in a case of tuberculous lymphadenopathy that mimicked malignancy of the pancreatic head and caused obstructive jaundice in an immunocompetent adolescent.

Objective: To detect the expression of Mycobacterium tuberculosis secreted protein Ag85B in paraffin-embedded tissues by immunohistochemistry (IHC), and to evaluate its application in the pathological diagnosis of tuberculosis., Methods: One hundred and five tuberculosis specimens (54 pulmonary tuberculosis, 51 lymph nodal tuberculosis) and 51 specimens of other diseases (8 lung cancer, 10 pulmonary abscess, 10 bronchiectasis, 7 lymphoma, 5 necrotizing lymphadenitis, 4 reactive hyperplasia lymphoid, and 7 sarcoidosis) were collected from January 2012 to July 2013 from Beijing Chest Hospital, Capital Medical University. One-step IHC was performed on paraffin-embedded tissues using antibody directed against Ag85B., Results: IHC and Ziehl-Neelsen (ZN) acid-fast staining showed that distribution and intensity of Ag85B expression were concordant with the distribution and number of acid-fast bacilli. IHC showed significantly higher sensitivity than ZN staining (50.5%, 53/105 vs. 31.4%, 33/105; χ² = 7.877, P = 0.005). The combined sensitivity of IHC and ZN staining was 59.0%. Moreover, oil immersion was not necessary for IHC, allowing more rapid diagnosis., Conclusion: IHC detection of Ag85B is a simple method with higher sensitivity than ZN staining, and demonstrated good value in the pathological diagnosis of tuberculosis.

CD8(+) T cells secreting Type1 and Type 17 cytokines and cytotoxic molecules play a major role in immunity and protection against pulmonary tuberculosis (PTB), although their role in tuberculous lymphadenitis (TBL) is not well known. To identify the distribution and function of CD8(+) T cells expressing Type1, Type 2 and Type 17 cytokines and cytotoxic molecules in TBL, we examined baseline and mycobacterial-antigen specific immune responses in the whole blood of individuals with PTB and compared them with TBL. TBL is characterized by elevated frequencies of baseline and mycobacterial-antigen stimulated CD8(+) T cells expressing Type 1 (IL-2 and TNFα) and Type 17 (IL-17A and IL-17F) cytokines in comparison to PTB individuals. In contrast, TBL individuals exhibited diminished frequency of CD8(+) T cells expressing perforin, granzyme B and CD107a. The blockade of IL-1R and IL-6R during antigenic stimulation resulted in significantly diminished frequencies of CD8(+) T cells expressing Type 1 and Type 17 cytokines in TBL. Therefore, our data suggest that TBL is characterized by an IL-1 and IL-6 dependent expansion of CD8(+) T cells expressing Type 1 and Type 17 cytokines as well as altered frequencies of cytotoxic molecules, reflecting an important association of these cells with the pathogenesis of TBL., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We present the case of a 51-year-old woman who presented a right supraclavicular mass corresponding to lymphadenitis with atypical mycobacterium (Mycobacterium genavense). Later, she developed subcutaneous purulent small nodules containing the same germ. The presence of this mycobacterium was in the context of recurrent peripheral T-cell lymphoma and immunosuppressive therapy for seronegative inflammatory polyarthritis.

Osteomyelitis due to Mycobacterium bovis Bacille Calmette-Guerin (BCG) often develops in patients with interferon-γ receptor 1 (IFNγR1) deficiency. In these patients, susceptibility appears to be caused by impaired interleukin-12- and IFNγ-mediated immunity. Here we report the case of a one-year-old girl with dominant partial IFNγR1 deficiency who suffered from lymphadenitis and multiple sites of osteomyelitis due to BCG infection. She was allergic to isoniazid and rifampicin--the prescribed standard treatment--and required prior desensitization therapy. She was subsequently treated with these drugs, but her symptoms did not improve. IFNγ therapy was added to the antitubercular therapy, increasing the serum level of IFNγ and leading to the resolution of the lymphadenitis and osteomyelitis. In conclusion, high dose IFNγ therapy in combination with antitubercular drugs led to resolution of BCG infection in a patient with dominant partial IFNγ deficiency., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Tuberculosis (TB) has recently re-emerged as a major public health threat worldwide. There is strong evidence that host genetic factors influence individual susceptibility to TB and that, once infected, young children and immunocompromised patients are at increased risk for mycobacterial disease and progression to extrapulmonary lymphadenitis., Methods: The association between polymorphisms of mannose-binding lectin and the susceptibility of 139 children with cervical mycobacterial lymphadenitis and infected with Mycobacterium tuberculosis was investigated., Results: The frequencies of genotypes A/B and B/B, based on codon 54 polymorphisms, were significantly different in TB-infected versus healthy control subjects. The frequency of the A/B genotype was significantly lower in TB-infected children (odds ratio = 0.56; 95% confidence interval: 0.36-0.87; P = 0.01), and the B/B genotype was significantly higher in TB-infected children (odds ratio = 4.68; 95% confidence interval: 1.35-16.3; P = 0.01) compared with healthy controls. The HYB haplotype appeared significantly more often to be protective in the healthy control population (odds ratio = 0.23; 95% confidence interval: 0.05-0.96; P = 0.046). Ex vitro phagocytosis assays indicated that high-expression mannose-binding lectin genotypes are associated with an increased risk of infection with M. tuberculosis., Conclusions: The present study suggests that mannose-binding lectin can protect against TB or predispose the host to the disease depending on the haplotype pair of the host. The low-expression genotype A/B and the HYB haplotype may be associated with protection against intracellular mycobacterial infections, whereas the high-expression genotype A/A may confer susceptibility to disease.

Forty-three patients with miliary tuberculosis were evaluated for diagnostic usefulness of enzyme-linked immunospot (ELISPOT) assay. Among noninvasive rapid tests available within 3-5 days, ELISPOT had the highest sensitivity (93%), compared with acid-fast bacilli stain (sputum, 32% and bronchoalveolar lavage, 7%), Mycobacterium tuberculosis polymerase chain reaction (sputum, 53% and bronchoalveolar lavage, 36%), and tuberculin skin test (22%). In comparison with 44 patients with lymph node tuberculosis, the sensitivity of the ELISPOT assay in patients with miliary tuberculosis (93%) was as high as in those with lymph node tuberculosis (95%, P = .63), whereas the sensitivity of the tuberculin skin test was substantially lower in patients with miliary tuberculosis (22%) than in those with lymph node tuberculosis (73%, P < .001).

Background: Th1 and Th17 responses are known to play an important role in immunity to pulmonary tuberculosis (PTB), although little is known about their role in extrapulmonary forms of tuberculosis (TB)., Methods: To identify the role of Th1, Th17, and Th22 cells in multi-focal TB lymphadenitis (TBL), we examined mycobacteria-specific immune responses in the whole blood of individuals with PTB (n = 20) and compared them with those with TBL (n = 25)., Results: Elevated frequencies of CD4(+) T cells expressing IFN- γ, TNF-α, and IL-2 were present in individuals with TBL compared with those with PTB at baseline and in response to ESAT-6 and CFP-10. Similarly, increased frequencies of CD4(+) T cells expressing IL-17A, IL-17F, and IFN-γ were also present in individuals with TBL at baseline and following ESAT-6 and CFP-10 stimulation although no significant difference in frequency of Th22 cells was observed. Finally, frequencies of Th1 (but not Th17) cells exhibited a significantly negative correlation with natural regulatory T cell frequencies at baseline., Conclusions: Multi-focal TB lymphadenitis is therefore characterized by elevated frequencies of Th1 and Th17 cells, indicating that Th1 and Th17 responses in TB disease are probably correlates of disease severity rather than of protective immunity.

Improved tools are required to study immunopathogenesis of tuberculosis (TB). Mycobacterium tuberculosis antigen-stimulated T cell-based assays can detect TB but are less effective when responses are compromised such as in severe disease. We investigated immune responses to M. tuberculosis whole sonicate (MTBs), recombinant antigens ESAT6 and CFP10 in whole blood cells of healthy endemic controls (EC, n = 42) and patients with pulmonary (PTB, n = 36) or extrapulmonary (ETB, n = 41) disease. Biomarkers of T cell activation (IFNγ) or modulation (IL10) and chemokines, CXCL9, CXCL10 and CCL2, secretion were measured. MTBs, ESAT6 and CFP10 all induced IFNγ responses in TB. ESAT6-induced IFNγ was elevated in TB as compared with EC. MTBs stimulated the highest IFNγ levels but did not differentiate between TB and EC. However, MTBs-induced CXCL10 (P = 0.004) was reduced, while IL10 (P < 0.001) was raised in TB as compared with EC. Between sites, MTBs-induced CCL2 (P = 0.001) and IL10 secretion was higher in PTB than ETB (P < 0.001). In comparison of disease severity, MTBs-induced IFNγ (P = 0.014) and CXCL10 (P = 0.022) levels were raised in moderate as compared with far advanced PTB. In ETB, MTBs-induced IL10 levels were greater in less-severe (L-ETB) than in severe disseminated (D-ETB) cases, P = 0.035. Within the L-ETB group, MTBs-induced IFNγ was greater in patients with tuberculous lymphadenitis than those with pleural TB (P = 0.002). As immune responses to MTBs were differentially activated in TB of different sites and severity, we propose the utility of MTBs-induced IFNγ, CXCL10 and IL10 as biomarkers in TB., (© 2011 The Authors. Scandinavian Journal of Immunology © 2011 Blackwell Publishing Ltd.)

The results of treatment of 12 patients, suffering complicated forms of abdominal tuberculosis and external intestinal fistulas, were presented. Late diagnosis of abdominal tuberculosis in the patients, suffering the complications phase of the disease, is caused by unclear symptoms presence in early stages of the disease. Clinical and laboratory indices in peritonitis of a phthisis origin are nonspeciphic. In 91% of patients, admitted to the hospital for complicated forms of abdominal tuberculosis and external intestinal fistulas, the operative treatment was indicated. Surgical intervention (more frequently right-sided hemicolectomy, enterostomy, the abscesses opening, the caseously-changed lymph nodes excision, formation of anastomosis) was performed in 11 patients for peritonitis and external intestinal fistulas. The method of a secure invagination anastomoses formation was elaborated, permitting to perform primary restoration operations. An early diagnosis, early effective therapy and radical surgical intervention conduction for complicated abdominal tuberculosis promote the patients to survive.

A 28-year-old woman was referred to our hospital for treatment of tuberculous lymphadenitis, after presenting with fever, left cervical lymphadenopathy, and a positive interferon-gamma release assay (QuantiFERON®-TB Gold In-Tube; QFT) result. Surprisingly, biopsy specimens of the cervical lymph nodes showed necrotic lesions with prominent nuclear debris and a proliferation of histiocytes, consistent with Kikuchi-Fujimoto disease (KFD). A diagnosis was made of KFD complicated by latent tuberculosis infection (LTBI), and all symptoms had resolved completely two months post-diagnosis. KFD may be misdiagnosed as tuberculous lymphadenitis, and antibiotics unnecessarily prescribed. Careful attention should therefore be paid when diagnosing cervical lymphadenopathy.

Background: Tuberculous lymphadenitis (TL) is the commonest form of extra-pulmonary tuberculosis in tropical countries., Objective: This study aimed to characterize in vivo and in vitro cellular immune responses to Mycobacterium PPD in TL patients as markers of disease and healing., Methods: Following informed consent, 36 TL patients, 40 patients with pulmonary tuberculosis (TB) and 20 apparently healthy individuals were enrolled when they met specific selection criteria. The tuberculin skin test (TST) and peripheral blood mono-nuclear cells (PBMCs) culture were conducted using PPD. The cytokines were measured using commercial kits., Results: The mean TST was 24.6 ±8.0 mm for TL patients. The TST was variable in pulmonary TB patients and healthy individuals. It was reactive in a third of pulmonary TB patients with a mean of 20 ±3.0 mm and reactive in half of the healthy individuals with a mean of 12.6 ±3.2 mm. Pre and post-treatment interferon gamma (IFN-g) mean levels were 498.6 ±905.8 pg/ml and 710.0 ±844.6 pg/ml respectively (p=0.0001) for TL patients, while IL-10 mean levels were 93.0 ±136.0 pg/ml and 32.4 ±31.7 pg/ml respectively (p= 0.0001). TST-reactive Pulmonary TB patients had significantly higher IFN-g (851 ±234.4 pg/ml) compared to TBLNT patients (p = 0.0001), while pulmonary TB patients had significantly lower IL-10 compared to TBLNT patients (p=0.0001). Apparently healthy individuals had significantly lower IFN-g and IL-10 levels compared to TBLNT and pulmonary TB patients (p=0.003)., Conclusion: Strong TST reactivity, high IFN-g and IL-10 levels are good surrogate markers of active TBLNT, while increasing IFN-g levels and decreasing IL-10 levels mark healing. Tuberculosis Skin Test reactivity although a good diagnostic marker does not disappear with treatment.

Background: In Sri Lanka, cutaneous leishmaniasis is a well-established disease caused by Leishmania donovani. Only a few cases of visceral disease and mucosal localization have been reported to date., Case Report: A 52-year-old man presented with severe local destruction of his upper and lower lip and total destruction of the anterior nasal septum and was diagnosed with mucosal leishmaniasis. The causative organism was confirmed to be Leishmania donovani. In addition he had tuberculous lymphadenitis and inherent immune deficiency. His previous medical history was unremarkable. The patient was successfully treated with intramuscular sodium stibogluconate., Conclusion: The clinical picture and satisfactory treatment response to antimony are similar to mucosal leishmaniasis caused by L. donovani reported in India and Sudan and with the absence of primary skin lesions make it different from new world mucosal leishmaniasis. Even though leishmania and tuberculous co-infection has been reported in association with HIV this has not been reported in inherent immune deficiency.

Objective: An increased risk of tuberculosis has been documented in humans treated with tumor necrosis factor alpha (TNFalpha)-neutralizing agents. In murine models, impaired signaling by TNF causes exacerbation of both acute and chronic infection associated with aberrant granuloma formation and maintenance. This study was undertaken to investigate immune modulation in the setting of TNF neutralization in primary and latent tuberculosis in a non-human primate model., Methods: Cynomolgus macaques 4 years of age or older were infected with Mycobacterium tuberculosis and subjected to clinical, microbiologic, immunologic, and radiographic examinations. Monkeys were classified as having active or latent disease 6-8 months after infection, based on clinical criteria. Monkeys used in acute infection studies were randomized to receive either adalimumab (prior to and during infection) or no treatment. Monkeys with latent infection that were randomized to receive TNF-neutralizing agent were given either an inhibitor of soluble TNF, recombinant methionyl human soluble TNF receptor I (p55-TNFRI), or adalimumab. Control monkeys with latent infection were given no treatment or saline. Data from previously studied monkeys with active or latent disease were also used for comparison., Results: Administration of TNF-neutralizing agents prior to M tuberculosis infection resulted in fulminant and disseminated disease by 8 weeks after infection. Neutralization of TNF in latently infected cynomolgus macaques caused reactivation in a majority of animals as determined by gross pathologic examination and bacterial burden. A spectrum of dissemination was noted, including extrapulmonary disease. Surprisingly, monkeys that developed primary and reactivation tuberculosis after TNF neutralization had similar granuloma structure and composition to that of control monkeys with active disease. TNF neutralization was associated with increased levels of interleukin-12, decreased levels of CCL4, increased chemokine receptor expression, and reduced mycobacteria-induced interferon-gamma production in blood but not in the affected mediastinal lymph nodes. Finally, the first signs of reactivation often occurred in thoracic lymph nodes., Conclusion: These findings have important clinical implications for determining the mechanism of TNF neutralization-related tuberculosis.

Background: Patients suspected of having cervical tuberculous lymphadenitis are diagnosed using investigations such as fine needle aspiration cytology and the polymerase chain reaction for Mycobacterium tuberculosis. However, these investigations are intended for primary tuberculosis infection. The majority of cervical tuberculous lymphadenitis cases in the elderly are thought to be caused by reactivation., Objective: The aims of this study were (1) to examine the efficacy of fine needle aspiration cytology, polymerase chain reaction and blood tests in the diagnosis of cervical tuberculous lymphadenitis caused by reactivation, and (2) to clarify any differences when compared with primarily infected cervical tuberculous lymphadenitis cases., Materials and Methods: Thirty-three elderly patients with neck lumps underwent excisional biopsy from 2003 to 2008. The efficacy of fine needle aspiration cytology was examined by comparing the results of excisional biopsy with those of fine needle aspiration cytology performed at the initial medical examination for cases of suspected tuberculous disease. Furthermore, the leucocyte count and C-reactive protein concentration were compared for cases of cervical tuberculous lymphadenitis versus cases of malignant lymphoma., Results: Although nine cases were diagnosed with cervical tuberculous lymphadenitis using excisional biopsy, only one of these had been suspected based on fine needle aspiration cytology results. Three cases with tuberculous lymphadenitis were suspected of having malignant lymphoma on initial examination. There was no significant difference in the leucocyte count and C-reactive protein concentration, comparing cases of tuberculous lymphadenitis versus malignant lymphoma., Conclusion: Unlike the primary infection often seen in endemic areas, the diagnosis of early stage tuberculous lymphadenitis of the swelling type caused by reactivation in elderly people is difficult to confirm unless excisional biopsy is performed. In elderly patients with neck lumps, cervical tuberculous lymphadenitis should be included in the differential diagnosis.

Bovine tuberculosis is a disease of worldwide importance yet comparatively little is known about chemokine responses to infection. We report on the levels of chemokine expression within lymph nodes of cattle infected with Mycobacterium bovis when infection would be well established. Expression levels of a number of chemokines were increased in infected cattle and could be correlated to levels of respective chemokine receptors. Several chemokines were significantly correlated to pathology within the lymph node, indicating a direct relationship between chemokine expression and disease. Vaccinated animals challenged with M. bovis had lower levels of chemokine expression than unvaccinated, challenged animals, correlating with lower levels of disease in vaccinated animals. The chemokine expression profile correlated with previous evidence for a pro-inflammatory bias within the lymph node. At this stage of infection we suggest there is on-going chemokine expression by cells associated with the granuloma and continual recruitment of cells to control infection.

Immune responses were assessed at the single-cell level in lymph nodes from children with tuberculous lymphadenitis. Tuberculosis infection was associated with tissue remodeling of lymph nodes as well as altered cellular composition. Granulomas were significantly enriched with CD68+ macrophages expressing the M. tuberculosis complex-specific protein antigen MPT64 and inducible nitric oxide synthase. There was a significant increase in CD8+ cytolytic T cells surrounding the granuloma; however, CD8+ T cells expressed low levels of the cytolytic and antimicrobial effector molecules perforin and granulysin in the granulomatous lesions. Quantitative real-time mRNA analysis revealed that interferon-gamma, tumor necrosis factor-alpha, and interleukin-17 were not up-regulated in infected lymph nodes, but there was a significant induction of both transforming growth factor-beta and interleukin-13. In addition, granulomas contained an increased number of CD4+FoxP3+ T cells co-expressing the immunoregulatory cytotoxic T-lymphocyte antigen-4 and glucocorticoid-induced tumor necrosis factor receptor molecules. Low numbers of CD8+ T cells in the lesions correlated with high levels of transforming growth factor-beta and FoxP3+ regulatory T cells, suggesting active immunosuppression at the local infection site. Compartmentalization and skewing of the immune response toward a regulatory phenotype may result in an uncoordinated effector T-cell response that reduces granule-mediated killing of M. tuberculosis-infected cells and subsequent disease control.

Genetic defects of interleukin (IL)-12/23-and interferon (IFN)-gamma-mediated immunity can cause increased susceptibility to intracellular microbes. Among these defects, a mutation of the gene encoding the IL-12 receptor beta1 (IL-12Rbeta1) is the most common worldwide. A 12-year old Thai boy with pre-existing neurofibromatosis type 1 (NF1) was evaluated for primary immunodeficiency after a history of tuberculous lymphadenitis, recurrent Salmonella infections and nocardiosis. Flow cytometry of phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs) revealed a defect in the IL-12Rbeta1 surface expression. A genetic study showed a novel nonsense homozygous mutation of the IL12RB1 gene in exon 4 (402C > A), confirming the diagnosis of IL-12Rbeta1 deficiency. This is the first case report of a primary IL-12Rbeta1 deficiency in Thailand with the interesting finding of a coexisting NF1.

During the pathogenesis of any granuloma, activated macrophages (AMs) are recruited and expanded under the influence of the migration inhibition factor. The goal of this study was to determine whether AMs then also proliferate by themselves or not. The proliferation index (PI) of AMs in lymph node biopsies from 40 cases of tuberculous lymphadenitis was evaluated by immunohistochemistry using a monoclonal antibody (MIB-1) to Ki-67. The PI was defined as the percentage of AMs with positive nuclear staining. It ranged from 3.6-20.6% (mean = 11.0 +/- 4.4) which was slightly lower than that reported in previous studies. Mitotic figures ranged from 0-6 per 500 AMs. Multinucleated giant cells ranged from 0-6.3 cells per low power field and their PI was exclusively 0%. Areas of caseous necrosis ranged from 5-85% of the total area of the tissue section examined (mean = 61.6 +/- 19.8). Mitotic figures, multinucleated giant cells, and areas of caseous necrosis lacked a statistically significant relationship to the PI (p > 0.05). In conclusion, AMs in granulomas can proliferate to a limited degree as detected by the mitotic figures and the PI.

The whole-blood interferon-gamma enzyme-linked immunosorbent assay (QuantiFERON-TB Gold [QFT-G]; Cellestis, Carnegie, Australia) has been studied mainly for diagnosing active pulmonary tuberculosis (TB) or latent TB. We prospectively evaluated its diagnostic usefulness in patients suspected with extrapulmonary TB (EP-TB). Of the 100 adult patients with suspected EP-TB, 43 were classified as "confirmed" EP-TB and 5 as "probable" EP-TB. Of the 48 with EP-TB, 27 (56%) were diagnosed with TB lymphadenitis and 11 (17%) with skeletal TB. The overall sensitivity and specificity of the assay were 69% (95% confidence interval [CI(95)], 53-81%) and 82% (CI(95), 69-91%), respectively. Among 44 patients presented with cervical lymphadenopathy, the QFT-G assay showed 86% (CI(95), 64-97%) sensitivity and 87% (CI(95), 66-97%) specificity, whereas in 28 with skeletal involvement, the sensitivity and specificity of the assay were 45% (CI(95), 17-77%) and 81% (CI(95), 54-96%), respectively. These suboptimal diagnostic performances suggest that the QFT-G assay alone is not sufficient for the diagnosis of EP-TB.

The data of a comprehensive study of 86 children aged 6 to 14 years, who were examined and treated at the Research Institute of Phthisiology for various manifestations of tuberculous infection: 25.6% with infected Mycobacterium tuberculosis with varying specific sensitization; 34.9% with minor forms of intrathoracic lymphatic tuberculosis (ITLT), 39.5% with disseminated processes into the intrathoracic lymph nodes, are analysed. Of the greatest informative value in the determination of the activity of tuberculous infection are RM V, VI, VII, and VIII dilutions in combination with immunological parameters of specific immunity: blast transpormation reaction (BTR) to PPD, a complex of serological reactions, IL-8, and lysosomal cationic test (LCT). Most children with ITLT showed a significant cellular response to PPD in the BTR test. It should be noted that on admission to the clinic, neutrophilic granulocytes were functionally inadequate in all the children as shown by LCT. The currently available immunological tests used in combination with the existing methods in the diagnosis of ITLT adequately evaluate the activity of tuberculous infection in children.

In modern society, there is a rise in the incidence of tuberculosis in all age groups, including children and adolescents. In old age group, a specific inflammation is detectable from Mantoux test results only in every four children. Tuberculous infection is diagnosed in half of cases when they turn to physicians for complains. Disseminated and complicated forms of tuberculosis are more frequently identified in these situations. The immune system has a particular emphasis on the course and outcome of the disease. The authors have established that caseous masses actively form, followed by the stimulation of the adequate cell pathway promoting the limitation of specific inflammation in old-age group children with primary tuberculosis. In secondary forms of tuberculous infection, there is an increase in the level of monocytes where the persistence and multiplication of the causative microorganism, as well as the activation of the humoral pathway inadequate for tuberculous infection are likely to occur, i.e. the infectious agent may be inhibited until activation of the Th-2 pathway of an immune response takes place.

Tuberculosis is the prototype of infections that require a cellular immune response for their control. It has been shown that CD4+ T-lymphocytes are most important in the protective response against Mycobacterium tuberculosis. CD8+ T-lymphocytes are also important for effective T-cell immune response. This study compares CD4+ and CD8+ baseline values in patients with different manifestations of tuberculosis. CD4+ and CD8+ in three groups of patients with tuberculosis (pulmonary, lymphadenitis, meningitis/milliary involvement) and a group of healthy volunteers were enumerated using flowcytometry. Twenty-six patients with pulmonary tuberculosis, 10 with adenitis, 16 with meningitis or milliary tuberculosis and 16 healthy volunteers entered the study. Mean CD4 in meningitis/milliary group was significantly lower than all other groups (p<0.05). Mean CD4 counts of patients with pulmonary tuberculosis was also significantly lower than control group (p=0.01). Mean CD8 in meningitis/milliary group was significantly lower than control group (p=0.02). No relation was found between results of TSTs and CD4 values in three groups. CD4 depletion is an expectable phenomenon in patients with tuberculosis. This study shows that patients with more severe form of disease had the lowest number of both CD4 and CD8 cells which can be a sign of suppressed cellular immunity in these patients.

Multi-colour flow cytometry was applied to determine T-cell-specific interferon-gamma, interleukin-2 and tumour necrosis factor-alpha expression in children with tuberculosis and non-tuberculosis mycobacterial lymphadenopathy (NTM-L). In vitro stimulation of peripheral blood mononuclear cells with purified protein derivative from Mycobacterium tuberculosis (tuberculin) and M. avium (sensitin) revealed differential recognition of tuberculin and sensitin in both study groups. Ratios of tuberculin-specific and sensitin-specific T-cell proportions in individual patients discriminated between children with tuberculosis or NTM-L. These findings have the potential to improve the differential diagnosis of mycobacterial infections.

Background: The immune response to Mycobacterium tuberculosis is complex and multifactorial, the cytokine system being a major factor in M. tuberculosis immunity., Aim: To analyze the immunohistochemical aspects of tuberculous lymph nodes in immunocompetent patients and search for associations between SOCS and cytokine expression in human tuberculous lymphadenitis., Methods: Thirteen lymph nodes were assayed by immunohistochemistry for SOCS-1 and 3, STAT-3, RANTES, MIP-1-alpha, ICAM-1, IFN-gamma as well as CD45RO, CD20, CD34, CD68, trypsin and lysozyme. Additionally, the RT in situ PCR was performed for SOCS-1 and 3 mRNA detection., Results: Decreased MIP-1 alpha expression together with reduced SOCS-3 (p=0.042), lysozyme (p=0.024) and CD45RO (p=0.05) was observed in the TB lymph nodes compared to the control lymph nodes. In conclusion, the lymphadenitis due to M. tuberculosis was associated with a downregulation of memory T cells (CD45RO), activated lysozymes and SOCS-3 compared to controls, which may play a role in the long-term bacterial replication and altered immune modulation characteristic of the disease.