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BACKGROUND: Neither the expression of CD14 and Toll-like receptor 4 (TLR4) on monocytes' surface nor the mutations CD14 -159TT and TLR4 Asp299Gly have yet been evaluated as risk factors for development of pulmonary tuberculosis (TB) in the Mexican population. METHODS: Level of membrane CD14 (mCD14) and membrane TLR4 (mTLR4) were determined by flow cytometry, in 104 patients with pulmonary TB (before and after treatment), 67 household contacts, and 114 healthy control subjects. Genotype/allele frequencies in CD14 -159 and TLR4 Asp299Gly were obtained by polymerase chain reaction-restriction-fragment length polymorphism. Levels of soluble CD14 (sCD14) in sera were quantified by ELISA. RESULTS: Higher levels of mCD14/sCD14 and mTLR4 were observed in the patients and the household contacts than in the control subjects (P<.05) and decreased in the patients after the infection was resolved. The frequency of the CD14 -159TT genotype was higher in the patients than in the control subjects (35.6% vs. 12.3%, respectively). Patients who were homozygous for allele T of the CD14 promoter gene had a significantly higher risk for development of pulmonary TB, with an odds ratio of 2.267 (95% confidence interval, 1.5%-3.3%). Levels of sCD14 or mCD14 were not associated with the CD14 -159TT genotype (P>.05).CONCLUSIONS: No association between TLR4 Asp299Gly and pulmonary TB was found. CD14 -159TT is a risk factor for development of pulmonary TB, whereas mCD14/sCD14 and mTLR4 are possible biomarkers for the prognosis for TB disease.CLINICAL TRIAL PROTOCOL ID: SA1168-05. [ABSTRACT FROM AUTHOR]

Pulmonary tuberculosis (PTB) and type 2 diabetes mellitus (T2DM) are two inflammatory diseases whose pathology involves neutrophils (NEU) as key participants. Countless inflammatory elements produced at the lesion sites leak into the blood and are distributed systemically. The study aimed to investigate the effect of the serum of patients with PTB, T2DM, and PTB + T2DM on the cellular and nuclear morphology of healthy NEU. Monolayers of NEU were prepared and incubated with sera from PTB (n꓿ 10), T2DM (n꓿10), PTB + T2DM (n꓿ 10) patients, or sera from healthy people (n = 10). Monolayers were stained for histones, elastase, and myeloperoxidase for NETosis, annexin V for apoptosis, and Iris fuchsia for necrosis. Hoechst stain (DNA) was used to identify the nuclear alterations. Necrosis was the predominant alteration. Sera from PTB + T2DM were the most potent change inducers. Normal sera did not induce cell alterations. The blood of TBP and T2DM patients carries a myriad of abnormal elements that induce necrosis of NEU in normal people, thus reflecting what might occur in the neutrophils of the patients themselves. These findings reinforce the participation of NEU in the pathology of these diseases. Necrosis is expected to be the most frequent neutrophil-induced alteration in tuberculosis and diabetes mellitus., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Is there a CD4+ and CD8+ immunity alteration in patients with pulmonary tuberculosis (TB) and diabetes (DM) that does not recover after antituberculosis treatment? This prospective comparative study evaluated CD4+ and CD8+ lymphocytic subpopulations and antituberculosis antibodies in patients with diabetes and tuberculosis (TB-DM), before and after antituberculosis treatment. CD4+ T cell counts were lower in patients with TB-DM compared to those with only TB or only DM, and these levels remained low even after two months of anti-TB treatment. Regarding the CD8+ T cell analysis, we identified higher blood values in the DM-only group, which may be explained by the high prevalence of latent tuberculosis (LTBI) in patients with DM. IgM antituberculosis antibodies levels were elevated in patients with only TB at baseline, and 2 months post-anti-TB treatment, IgG did not express any relevant alterations. Our results suggest an alteration in CD4+ immunity in patients with TB-DM that did not normalize after antituberculosis treatment.

Neutrophils are the most abundant leukocytes in the bloodstream and are very important for the resolution of infection. One of the strategies used by neutrophils to eliminate a microorganism is the formation of extracellular traps. Different methods for neutrophil extracellular traps (NETs) visualization have been described along the years, usually requiring the use of a fluorescent, confocal or scanning electron microscope. This research aimed to visualize NETs using light microscopy as another way to study NETs prior to using the more expensive techniques, making NETs research more cost effective. We evaluated neutrophil purity, viability and function by analyzing the formation of NETs comparing DAPI with safranin. When evaluating NETs formation, neutrophils that were not stimulated did not form NETs and when neutrophils were exposed to PMA or S. aureus NETs were formed and visualized with safranin under light microscopy and DAPI under fluorescence microscopy. Our method demonstrates another way to visualize NETs that can be added to the standard methods of visualization of NETs, increasing the opportunities to generate knowledge in the topic in any lab around the world., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors. Published by Elsevier B.V.)

Background: It has been reported that sera from patients with active pulmonary tuberculosis (APT) induced nuclear changes in normal neutrophils that included pyknosis, swelling, apoptosis, and production of extracellular traps (NETs). Similar changes were observed with some sera from their household contacts but not with sera from healthy, unrelated individuals. It was suggested that those sera from household contacts that induced neutrophil nuclear changes might correspond to people with subclinical tuberculosis. Thus, our experimental approach might serve to identify individuals with early, ongoing disease., Methods: Nuclear changes in neutrophils were fully evident by 3 h of contact and beyond. Circulating mycobacterial antigens were the most likely candidates for this effect. We wanted to know whether the nuclear changes induced on neutrophils by the sera of APT patients would negatively affect the phagocytic/microbicidal ability of neutrophils exposed to APT sera for short periods., Results: We now provide evidence that short-term contact (30 min) with sera from patients with pulmonary tuberculosis increases several phagocytic parameters of normal neutrophils, including endocytosis, myeloperoxidase levels, production of free reactive oxygen species, phagolysosome fusion, and microbicidal activity on Staphylococcus aureus, with these effects not being observed with sera from healthy donors. We also give evidence that suggests that ESAT-6 and CFP-10 are involved in the phenomenon., Conclusion: We conclude that activation is a stage that precedes lethal nuclear changes in neutrophils and suggests that autologous neutrophils must circulate in an altered state in the APT patients, thus contributing to the pathology of the disease., Competing Interests: None

Strongyloides stercoralis hyperinfection syndrome is a medical emergency that requires a high level of suspicion. Immunocompromised patients are at high risk of hyperinfection syndrome; however, malnutrition, alcoholism, and diabetes mellitus also need to be considered as predisposing factors. The diagnosis and treatment of Strongyloides hyperinfection are challenging and patients often have severe complications. Consequently, mortality is overwhelmingly high, with proportions above 60%. Herein, we report a case of Strongyloides hyperinfection in a 40-year-old alcoholic diabetic patient living in México. Unfortunately, the late diagnosis resulted in his death despite the treatment and supportive measures. Increased awareness is needed to prevent the dire consequences of strongyloidiasis.

Encephalitozoon cuniculi is a fungi-related, obligate, zoonotic, spore-forming intracellular eukaryotic microorganism. This emerging pathogen causes granulomas to form in the brain and kidneys of infected individuals. The objective of the current study was to detect the distribution of TNF-α- and IL-4-positive cells using immunohistochemistry within these granulomas in both infected immunocompetent (group A) and immunosuppressed (group B) New Zealand white rabbits. In the brain, labeled TNF-α immune cells were mainly located in the granuloma peripheries in group B. Granulomas examined in the kidneys of groups A and B were TNF-α positive, but were significantly different (p < 0.001) when compared with the brain. IL-4-producing immune cells in the brain and kidneys were disseminated within granulomas in groups A and B; however, no significant difference (p > 0.05), was observed. IL-4 positive cells were more numerous in brain sections of group B and differed significantly (p < 0.05) when compared with kidneys. Granulomas were not observed in control animals (groups C and D). In conclusion, we identified TNF-α positive cells in both the brain and kidneys of immunocompetent and immunosuppressed animals; IL-4 positive cells were numerous in the brains of immunosuppressed rabbits; however, in terms of percentage were numerous in the brains of immunocompetent rabbits. Immunosuppression appeared to stimulate a change in the cellular phenotype of Th1- to Th2-like granulomas in the brain and kidneys via an unknown mechanism. Expression of pro- and pre-inflammatory cytokines in microsporidian granulomas suggests a mechanism by which E. cuniculi evades the immune response, causing more severe disease. These results increase our understanding of TNF-α and IL-4-positive cells within the E. cuniculi granuloma microenvironment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Encephalitozoon cuniculi (E. cuniculi) is a fungi-related, obligate, zoonotic, spore-forming intracellular eukaryotic microorganism. This emerging pathogen causes granulomas in brain and kidneys of infected individuals. The objective of this study was to detect the distribution of CD4, CD8 and MHCII-positive cells within granulomas in these organs in infected immunocompetent (group A) and infected immunosuppressed (group B) New Zealand white rabbits using immunohistochemistry. In brain, labeled CD4 immune cells were mainly located in the periphery of granulomas in group B. Kidneys of groups A and B, displayed CD4-positive in granulomas and were significant different when compared to brain. CD8 immune cells in brain and kidneys were disseminated in the granulomas in groups A and B; however, no significant difference was observed. MHCII-positive cells were more numerous in brain sections of group B and were significantly different when compared to kidney sections. Granulomas were not observed in control animals of group C and D. In conclusion, we identified CD4-positive cells in both the brain and kidneys of immunocompetent and immunosuppressed animals; CD8-positive cells were more numerous in brain of immunosuppressed rabbits and MHCII cells were more predominant in brain of immunocompetent rabbits. Apparently, the immunosuppression stimulated a change in the cellular phenotype of Th1- to Th2-like granulomas in brain and kidneys by an unknown mechanism. These results increase our understanding of CD4, CD8 and MHCII positive cells within the E. cuniculi granuloma microenvironment and will help in future microsporidian granulomas studies of both immunocompetent and immunosuppressed individuals., Competing Interests: Declaration of Competing Interest The authors declared no conflicts of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Opportunistic parasites are still important agents causing morbidity and mortality in immunocompromised patients, particularly those living with HIV/AIDS. Few studies in Mexico have attempted to determine the prevalence of opportunistic intestinal parasites causing diarrhea in immunocompromised patients. A study was conducted to determine the intestinal parasites in HIV-positive and HIV-negative immunocompromised patients with diarrhea admitted to a tertiary care hospital in Monterrey, Mexico, from 2014 to 2015. Stool samples were examined for trophozoites, cysts, and eggs using the EGRoPe sedimentation-concentration technique and special techniques (modified Ziehl-Neelsen stain, modified trichrome stain). A total of 56 patients were included. The overall prevalence of intestinal parasitism was 64% (36/56); 22/36 patients were HIV-positive. Prevalence of opportunistic parasites was 69% in HIV-infected patients compared to 44% in HIV-negative patients (P = 0.06). Microsporidia were the most frequently identified parasites (24/36, 67%), followed by Cryptosporidium sp. (6/36, 17%), Sarcocystis sp. (4/36, 11%), Cystoisospora belli (3/36, 8%), and Cyclospora cayetanensis (1/36, 3%). Overall prevalence rates of microsporidiosis and cryptosporidiosis were 43% and 11%, respectively. Among HIV-infected patients, prevalence rates of microsporidiosis and cryptosporidiosis were 48% and 14%, respectively. We also report the first cases of intestinal sarcocystosis in Mexico, all in HIV-infected patients. In conclusion, microsporidia and coccidia are major parasitic agents causing diarrhea in immunocompromised patients, particularly HIV-infected patients.

Coccidioidomycosis is a fungal disease caused by Coccidioides immitis or Coccidioides posadasii. These fungi are endemic in the southern USA and northern Mexico. Immunocompromised patients are susceptible to develop severe forms of this fungal infection. Cytokines play an important role in controlling the fungal infection, but little is known about the predominant immunological environment in human lung tissue from fatal cases. Our aim was to analyze the pro-inflammatory and anti-inflammatory cytokines and monocyte/macrophages markers (CD14 and CD206) in the granulomas of six fatal cases of coccidioidomycosis. Cytokines and surface markers were higher in coccidioidomycosis cases when compared to control (P < 0.05). CD14 positive cells were increased inside the coccidioidal granuloma when compared to the outside (P < 0.05). No differences were found in the number of CD206+ cells inside the granuloma when compared to the outer population (P > 0.05). Interestingly, an analysis of stain intensity signals showed an increased signaling of CD14, CD206, IL-10 and TNFα inside the granuloma when compared to the outside (P < 0.05). iNOS and IL-12 gene expression were not detected in coccidioidomycosis cases, while IL-10, IL-6 and TGFβ gene expression were detected, but the differences when compared to healthy lungs were not significant (P > 0.05). TNFα gene expression was lower in coccidioidomycosis cases when compared to healthy lung (P = 0.05). In conclusion, pro- and anti-inflammatory responses co-exist inside of the granulomas of fatal cases of coccidioidomycosis and the absent of iNOS and IL-12 gene expression may be related with patient's outcome.

Levels of interferon (IFN)-γ and interleukin (IL)-10 were measured in the serum of immunocompetent and immunosuppressed New Zealand White rabbits naturally infected with Encephalitozoon cuniculi. IFN-γ levels were elevated in infected rabbits, and a synergic effect was observed in animals treated with the immunosuppressive agent dexamethasone (Dex). The role of IL-10 in infected rabbits remains unclear, as IL-10 levels were similar to those of negative controls. Dex appeared to exhibit a proinflammatory effect, as IFN-γ levels were elevated in infected immunosuppressed rabbits. Similarly, Dex exhibited a synergic effect in infected immunosuppressed rabbits, as evidenced by the elevation in IFN-γ production. These data indicate that the immune response to this glucocorticoid should be considered in the design of future animal model studies of immunosuppression., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Human P-glycoprotein (P-gp) is a membrane transporter encoded by ABCB1 (also known as MDR1) that plays a critical role in pharmacokinetics of many unrelated drugs. Rifampin (RMP) and ethambutol (ETB), two anti-tubercular agents, are substrates of P-gp. Single nucleotide polymorphisms (SNPs) in ABCB1 have been associated with resistance to several drugs; however, their association with RMP and ETB resistance in tuberculosis patients has not yet been studied. Genotype/allele frequencies in C1236T, G2677T/A and C3435T SNPs of ABCB1 were obtained from 99 tuberculosis patients susceptible or resistant to RMP and ETB (NoRER or RER). 2677G>A allele prevalence was found to be significantly higher in the RER group compared to NoRER (5 resistant vs 2 non-resistant patients, P < 0.01; OR, 11.0; 95% CI, 2.00-56.00). No differences were found in genotype/allele frequencies in C1236T and C3435T SNPs of ABCB1 and resistance to RMP and ETB in tuberculosis patients (P > 0.05). The present study suggests the 2677G>A allele of ABCB1 could be associated with simultaneous resistance to RMP and ETB in pulmonary tuberculosis patients. Further studies with larger sample sizes are needed to confirm this association and explore its nature., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Although current polyvalent vaccines can prevent development of cervical cancer, they cannot be used to treat patients who already have the disease. Adenovirus expressing calreticulin-E7 (Ad-CRT-E7) has shown promising results in the cervical cancer murine model. We also demonstrated that immunization with Lactococcus lactis encoding HPV-16 E7 (Ll-E7) anchored to its surface induces significant HPV-16 E7-specific immune response. Here, we assessed the combination of both approaches in the treatment of a cervical cancer animal model. Intranasal preimmunization of Ll-E7, followed by a single Ad-CRT/E7 application, induced ∼80% of tumor suppression in comparison with controls. Mice treated with a combination of Ll-E7 and Ad-CRT/E7 resulted in a 70% survival rate 300 days post-treatment, whereas 100% of the mice in the control groups died by 50 days. Significant CD8+ cytotoxic T-lymphocytes infiltration was detected in the tumors of mice treated with Ll-E7+Ad-CRT/E7. Tumors with regression showed a greater number of positive cells for in situ TUNEL staining than controls. Our results suggest that preimmunization with Ll-E7 enhances the Ad-CRT/E7-mediated antitumor effect. This treatment provides an enormous advantage over repeated applications of Ad-CRT/E7 by maintaining the effectiveness of the three-dose application of Ad-CRT/E7, but avoiding the high systemic toxicities associated with such repeat treatments.

Chemokines are members of the super family of cytokines necessary for leukocyte recruitment in tissues and lymphoid organs. The interferon-gamma inducible protein-10 (IP-10) chemo-attracts CXCR3-expressing cells, such as activated T lymphocytes and monocytes. We have genetically engineered a strain of Lactococcus lactis to secrete a biologically active murine IP-10 that interacts with human CXCR3, its homolog receptor, and chemo-attracts human CD3+ T lymphocytes.

CD11c is involved in Mycobacterium tuberculosis phagocytosis by human macrophages. CD11c has not been evaluated during antituberculosis (anti-TB) treatment. CD11c expression was evaluated on monocytes from peripheral blood leukocytes of pulmonary TB (PTB) patients and healthy controls by flow cytometry, whereas CD11c/Arg47Trp polymorphism was analyzed using polymerase chain reaction-restriction fragment length polymorphism. PTB patients had increased levels of CD11c on blood monocytes as compared to healthy controls. CD11c levels decreased in response to anti-TB treatment. CD11c/Arg47Trp polymorphism is not associated with PTB.

Tuberculosis is the most frequent coinfection in humans infected with HIV-1, but little is known about mechanisms that favors coinfection. The aim of this work is to understand tuberculosis and HIV infections. We determined the pattern of expression of CD11c, CD14, CD40, CCR5, and CXCR4 and quantified IL-1beta, IL-6, IL-8, TNF-alpha, and RANTES in tuberculosis patients and HIV patients. Monocytes from healthy PPD+ volunteers (HP(+)V) stimulated with intracellular proteins (IP), lipids, and polysaccharides (PLS) from Mycobacterium tuberculosis down regulate CD11c expression (p < 0.05). On the contrary, CD14 expression was elevated in tuberculosis patients (p < 0.05) and HIV-infected patients (p > 0.05). CD14 expression was elevated on monocytes from HP(+)V stimulated with PLS and lipids (p < 0.05). CD40 low expression was found in tuberculosis patients and on monocytes from HP(+)V stimulated with lipids, but it was elevated in HIV-infected patients (p < 0.05). CXCR4 and CCR5 expression was high in pulmonary tuberculosis patients and low in HIV-infected patients (p < 0.05). Finally, CCR5+ monocytes from HP(+)V after stimulation with PLS and CXCR4+ lymphocytes were elevated after stimulation with IP (p < 0.05). In general, high levels of IL-1beta, IL-6, and TNF-alpha were found in all groups, but low levels of RANTES were found in pulmonary tuberculosis patients. In conclusion, the pulmonary tuberculosis patients have a microenvironment that facilitates the HIV infection through three possible mechanisms: (1) increasing the coreceptor for HIV entrance, (2) increasing proinflammatory cytokines, and (3) down-regulating RANTES. At the same time, HIV patients have a microenvironment that facilitates entry of M. tuberculosis into macrophages through CD14.

Background: Individuals with Down's syndrome (DS) differ in their oral condition compared with the healthy population. Periodontal disease in persons with DS under the age of 30 years is very high. Immune deficiencies are also present. For dental practitioners it is difficult to decide on a particular course of treatment. In this study, patients with DS were selected in order to 1) evaluate the effectiveness of surgical and non-surgical periodontal therapies and 2) assess their immunological status., Methods: The population consisted of 14 DS patients (14 to 30 years old). Surgical and non-surgical periodontal therapies were compared in a split-mouth design. Clinical measurements of plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment levels (CAL) were taken at baseline, posttreatment, 6 months, and 1 year. Immunomodulatory activity of neutrophils was analyzed in vitro by chemotaxis (Boyden migration chamber), phagocytic activity, and production of super-oxide anion (NBT reduction) tests and compared between DS patients and healthy controls., Results: Both surgical and non-surgical therapies showed a significant improvement in all the clinical parameters compared to baseline. There were no differences between surgical and nonsurgical therapy in PI or GI. There was a significant PD reduction with the non-surgical therapy at 1 to 3 mm PD. However in PD >3 mm the surgical therapy, although not statistically significant, showed better results. Neutrophil chemotaxis, phagocytic activity, and production of super-oxide anion were significantly decreased in the DS patients., Conclusions: After a year, both surgical and non-surgical therapies have similar periodontal clinical improvement in DS patients. There is partial impairment of immunological functions in DS individuals which does not seem to affect the clinical response to therapy.