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Introduction. Coccidiosis, caused by protozoan parasites of genus Eimeria, is a disease with large impact on poultry production worldwide. It is well known that Eimeria immunity is dependent on Th1-type responses. Gap Statement. In vitro assessment of Eimeria-specific T-cell activity would therefore be a valuable research tool but has so far proven difficult to establish. Aim. The present study aimed to evaluate in vitro induced blast transformation and CD25 expression in defined chicken T-cell populations as a measure of Eimeria immunity. Methodology. Three E. tenella infection experiments were performed and PBMC and/or spleen cells were collected between 6 and 16 days after infection of chickens. Cells were stimulated in vitro with E. tenella antigens and T-cell activation was assessed by immunofluorescence labelling and flow cytometry. Results. The results consistently showed statistically significant E. tenella specific activation of TCRα/β+T cells within a ‘window’ from 8 to 14 days after infection for both spleen cells and PBMC. Responding T-cells were identified as CD4+CD8-, CD4+CD8αα+ and CD4-CD8αβ+ where the CD4+CD8αα+ cells generally showed the highest responses. All three of these TCRα/βT-cell subsets showed significant E. tenella induced blast transformation and/or CD25 expression albeit not always in concert on the same days after infection indicating complex kinetics of T-cell responses. In general, responses were higher for spleen cells compared to PBMC for all responding T-cell populations. Conclusions. This methodology shows promise to study Eimeria-specific T-cells, e.g. to evaluate vaccine responses. Results indicated that a Th1-type response was induced and suggested a role for CD4+CD8αα+ cells in Eimeria immunity.

Bone marrow fibrosis leading to dry tap aspiration and often associated with blast crisis has previously been reported in both Chronic myeloid leukemia and Primary myelofibrosis. The similarities between these two conditions in terms of clinical presentation and morphology can really create a diagnostic dilemma. Here we present a case of Chronic myeloid leukemia in fibrosis and blast crisis in a 32 year old lady which closely resembled Primary myelofibrosis in transformation. All myeloproliferative neoplasms can undergo blast transformation. In this case, the detection of Philadelphia chromosome helped to distinguish Chronic myeloid leukemia from other myeloproliferative neoplasms.

Case report - Introduction We present a rare case of a patient with a culture-negative mono arthritis. Our patient had a history of Essential Thrombocythemia (ET), and his mono arthritis initially responded to antibiotics. He had a fluctuating clinical course, and a multi-disciplinary approach with rheumatology, orthopaedics, haematology and radiology was needed. After multiple MRIs and two bone biopsies, he was diagnosed with acute myeloid leukaemia (AML) with blast transformation of his myeloproliferative neoplasm, essential thrombocythemia. This case highlights the need to consider the differentials of mono arthritis despite initial response to treatment, and the value of a multi-disciplinary approach. Case report - Case description A 59-year-old male was referred to rheumatology with new right hip pain, and high inflammatory markers. He was negative for infection, trauma, seronegative or crystal arthritis. Past history of ET diagnosed in 2011 with high-risk genetics. At haematology review 3 months prior he had new pancytopenia with deranged LFTs. A bone marrow biopsy (BMBx) showed post-ET myelofibrosis with no circulating blasts. His hydroxycarbamide and aspirin were stopped and blood counts recovered without other intervention. He was afebrile and had a right hip effusion. The hip aspirate revealed no growth after 48 hours and negative crystals. Bloods: Hb 101 WCC 5.1 Neutrophils 3.8 Platelets 318 CRP 276 ESR 72 Urate normal. MRI showed a right hip joint effusion, with extensive soft tissue oedema likely secondary to infection. He was commenced on IV Flucloxacillin, clinically improved and was discharged to complete antibiotics. At review in 2 weeks he had worsening hip pain, night sweats, raised CRP, and was anaemic with normal WCC and platelets. A repeat MRI showed evolving abscess formation within the acetabular roof. After discussion with haematology and orthopaedics, the clinical impression was septic arthritis as he was responding to treatment, his blood film was normal and recent BMBx showed no transformation. The plan was to continue with antibiotics and to perform a bone biopsy. Bone biopsy showed necrotic debris secondary to the underlying osteomyelitis. Due to fluctuating course a repeat MRI was performed showing a larger hip effusion with appearances suspicious for malignancy. A repeat bone biopsy showed post ET fibrosis with blasts and large tumour cells. A blood film showed peripheral blasts circulating. The unifying diagnosis was AML with blast transformation of myeloproliferative neoplasm. Our patient was refractory to chemotherapy. He was treated with palliative radiotherapy and died with palliative care input. Case report - Discussion Septic arthritis typically affects one joint and mimics include abscess, AVN, cellulitis, crystal, malignancy, reactive and inflammatory arthritis, and transient synovitis. Synovial fluid analysis is the gold standard for the diagnosis and will demonstrate growth in approximately 80% of all cases of non-gonococcal septic arthritis. The remaining 20% are ‘culture negative’ and may demonstrate no growth for a variety of reasons. 14% of patients with ‘culture negative’ septic arthritis will have positive blood cultures and newer diagnostic tools such as PCR testing of synovial fluid increases the likelihood of correctly identifying an organism. Our patient initially responded to treatment for septic arthritis; however, the atypical clinical course and history of ET prompted further investigations for differentials of ‘seronegative arthritis.’ ET is a myeloproliferative neoplasm with a prevalence of 40/100000. It is usually indolent and recognised genes are JAK2, CALR and MPL. Complications include vascular events such as thrombosis and haemorrhage. More serious complications of transformation to myelodysplasia, AML or myelofibrosis are rare with an incidence of 1—5% and median time to transformation is between 84 and138 months. Rheumatic manifestations of leukaemia include leukaemia arthritis. This is uncommon and develops due to infiltration of synovial membrane by leukaemia cells. Synovial fluid or synovial biopsy can be analysed for blasts and flow cytometry. The reported incidence is 12% in children, and only case reports in adults. The blast infiltrative phase of leukaemia usually occurs in the bone marrow, or blood. Previous case reports describe extra medullary blasts present in lymphoid tissues, and here we report an extra medullary blast sarcoma. Transformation of ET to AML is typically very difficult to manage, with a poor prognosis. Treatment options include chemotherapy, stem cell transplants and the JAK inhibitor ruxolitinib. Case report - Key learning points Differentials for a mono arthritis are wide and this is a rare case of hip mono arthritis as the clinical presentation of transformation of an ET to AML. Septic arthritis is one of the most important diagnoses to rule out in a patient with a mono arthritis and high inflammatory markers. Up to 20% of septic arthritis can be ‘culture negative.’ Peripheral blood cultures and PCR testing of synovial fluid can be used to enhance the identification of organisms. Although our index of suspicion was high, our patient was not displaying typical manifestations of transformation to AML. The development of an acute leukaemia can be insidious, and case reports have described infiltration of bone marrow simultaneously or months following an extra medullary blast crisis. This case highlights the importance of further, and if necessary repeated, investigations in a patient with an atypical history or clinical course. Malignancy as a differential for mono arthritis is important to consider as mis diagnosis could delay treatment and prognosis is very different. Both bone biopsies yielded abnormal bone; however, it was the second biopsy that showed circulating blasts. In our case the bone biopsies were performed without radiological guidance, and studies have shown that CT guided biopsies have a higher yield. In future, consideration could be given to utilising radiologically guided biopsy. A multi-disciplinary approach between a number of specialities is often needed in complex cases. This case highlights the value of this with collaboration with our haematology, orthopaedic, radiology and pathology colleagues.

Chronic Myelogenous Leukemia (CML) is a chronic myeloproliferative neoplasm characterised by uncontrolled proliferation of mature and maturating granulocytes. In our case 42 year male over a period of 13 months showed blastic transformation with severe thrombocytopenia, disseminated intravascular coagulation and bleeding. Herewith we present this case for its clinical behaviour, hematological findings and disease course. Keyword: Myeloproliferative Neoplasms, Blast crisis, Leukemia.

Extracts from aerial parts of G. aparine (cleavers) constitute a herbal remedy with monography in British Herbal Pharmacopeia. On the European market, there are several drugs and food supplements consisting of Galium extracts. In folk medicine, cleavers was used topically in Europe, Asia, and the Americas to treat skin diseases. In several remedies, cleavers is also listed as an immunomodulatory active herb influencing the defense response of the human body. The aim of this study was to investigate the immunostimulatory activity and antioxidant potential in vitro of a raw infusion of cleavers and bioactive fractions. The functional activity of lymphocytes in the reaction of the lymphocyte blast transformation (RLBT) method was used for immunomodulatory activity assays and direct scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and hydrogen peroxide (H2O2) was chosen for the examination of antioxidant activity. It was shown that both the raw extract and fractions show significant immunostimulatory and scavenging activities. The obtained data partially justify the traditional use of cleavers as topical remedy for skin infections and for wounds.

The phenomenon of population aging has been observed in many countries in recent years due to the increase in human life expectancy. Therefore, studies of lymphoid tissue, which supports the immune processes in the human body, are now of particular importance. The lymphoid tissue of appendices from humans of age groups of 54–81 years and 26–35 years was histologically studied, and the results were statistically treated. The lymphoid tissue belonging to senile age groups showed reductions and rearrangements with partial integrity of the modified lymphoid nodules. The majority of nodules were without lighter-stained germinal centers, and the processes of blast transformation were preserved therein. Mitoses were observed in the central zones of nodules with lighter centers. Lymphocytes actively migrate to lymphatic capillaries. The condition of lymphoid tissue in the appendix depends on the individual body parameters. In some cases, the lymphoid tissue is completely reduced, and the organ’s lumen is closed.

Introduction: There are no standard salvage options for patients (pts) with chronic myelomonocytic leukemia (CMML) after progressing on hypomethylating agents (HMA). Outcomes are especially poor after blast transformation (BT). Combination therapy with the BCL2 inhibitor venetoclax (VEN) has emerged as an efficacious treatment for pts with AML. To the best of our knowledge, the outcomes of VEN based therapies specifically in CMML, with or without BT, have not been reported. Methods: We retrospectively reviewed pts with CMML, both with and without BT, treated with VEN-based therapy at our institution from 2016 to present. Mutation testing at baseline (N=31) and loss of response was performed using an amplicon-based next-generation sequencing panel. Pts were stratified according to CPSS-Mol and whether VEN was given for chronic vs. transformed disease. Responses for CMML and CMML-BT were defined by IWG 2006 and ELN 2017 criteria, respectively. Cox proportional hazard regression and the Kaplan-Meir product limit method were used to study association of variables with survival. Overall survival was defined as the time from start of VEN therapy to death or last follow up and was not censored at date of transplant. Results: Thirty-two pts were identified: 17 CMML, 15 CMML-BT. Pt characteristics are shown in Figure 1A. The median age was 70 years in both groups; most pts were men with a PS of 0-1. The majority of pts treated for CMML had proliferative subtype (53%) and CPSS-Mol intermediate-2/high (76%) risk disease at diagnosis. Both groups were enriched for diploid karyotype (53 and 60%). Frequency of mutations prior VEN-based therapy are shown in Figure 1B; TET2 (71%), ASXL1 (55%), RUNX1 (45%), and NRAS (39%) were the most common. Forty-one percent (CMML) and 67% (BT) of pts had received ≥2 previous lines of therapy, and most (88% CMML, 87% BT) received VEN following HMA failure. The median per-cycle dosing of VEN was 100 mg x 14 days (CMML) and 200 mg x 21 days (BT). VEN was given in combination with HMA in 76% of the CMML group, whereas in the BT group it was partnered with HMA (47%), intensive chemotherapy (20%), or other agents (33%). Almost all pts were concurrently on a triazole antifungal. Forty-seven percent of pts in both groups required admission to hospital for infection and/or fever while on VEN. After a median of 1 (range 1-6 CMML, 1-16 BT) cycle in both groups, ORR was 59% (CR 0%, mCR 53%, PR 6%) in the CMML and 67% (CR/CRi 53%, MLFS 7%, PR 7%) in the BT group. Five out of the 7 pts in the BT group with CR/CRi responses and evaluable MRD by flow cytometry (sensitivity 0.01%) were MRD negative. Presence of a RAS pathway mutation was associated with lower likelihood of achieving CR (17% vs 83%, OR 5.5, 95% CI 0.92-33.2, p Conclusions: Combination therapy with venetoclax has activity in both CMML and secondary AML arising out of CMML, even in the challenging setting of relapsed/refractory disease post HMA failure. It appears particularly useful as a means to clear excess marrow blasts and to serve as a bridge to allogeneic SCT. Without a subsequent consolidative strategy, however, response durations are short. RAS pathway mutations were associated with lower likelihood of CR and often emerge at relapse. NPM1 mutations predict for CR and RUNX1 mutations are associated with a trend to higher ORR. The high incidence of neutropenic infections in this cohort suggests that venetoclax dosing should be carefully tailored to minimize myelosuppression. Figure 1 Disclosures DiNardo: ImmuneOnc: Honoraria; Calithera: Research Funding; Novartis: Consultancy; Celgene: Consultancy, Honoraria, Research Funding; Notable Labs: Membership on an entity's Board of Directors or advisory committees; Agios: Consultancy, Honoraria, Research Funding; AbbVie: Consultancy, Honoraria, Research Funding; MedImmune: Honoraria; Takeda: Honoraria; Syros: Honoraria; Jazz: Honoraria; Daiichi Sankyo: Consultancy, Honoraria, Research Funding. Konopleva:Amgen: Consultancy; Calithera: Research Funding; Rafael Pharmaceutical: Research Funding; Sanofi: Research Funding; Agios: Research Funding; Reata Pharmaceutical Inc.;: Patents & Royalties: patents and royalties with patent US 7,795,305 B2 on CDDO-compounds and combination therapies, licensed to Reata Pharmaceutical; Stemline Therapeutics: Consultancy, Research Funding; F. Hoffmann La-Roche: Consultancy, Research Funding; Kisoji: Consultancy; Cellectis: Research Funding; AbbVie: Consultancy, Research Funding; Genentech: Consultancy, Research Funding; Ablynx: Research Funding; Forty-Seven: Consultancy, Research Funding; AstraZeneca: Research Funding; Ascentage: Research Funding; Eli Lilly: Research Funding. Borthakur:Treadwell Therapeutics: Consultancy; Polaris: Research Funding; Xbiotech USA: Research Funding; Nkarta Therapeutics: Consultancy; BioTherix: Consultancy; BioLine Rx: Consultancy; PTC Therapeutics: Consultancy; Argenx: Consultancy; FTC Therapeutics: Consultancy; Curio Science LLC: Consultancy; Oncoceutics: Research Funding; BioLine Rx: Research Funding; Cyclacel: Research Funding; GSK: Research Funding; Jannsen: Research Funding; Abbvie: Research Funding; Novartis: Research Funding; Incyte: Research Funding; PTC Therapeutics: Research Funding; AstraZeneca: Research Funding; BMS: Research Funding. Sasaki:Otsuka: Honoraria; Daiichi Sankyo: Consultancy; Novartis: Consultancy, Research Funding; Pfizer Japan: Consultancy. Jabbour:Genentech: Other: Advisory role, Research Funding; Amgen: Other: Advisory role, Research Funding; Pfizer: Other: Advisory role, Research Funding; BMS: Other: Advisory role, Research Funding; Takeda: Other: Advisory role, Research Funding; AbbVie: Other: Advisory role, Research Funding; Adaptive Biotechnologies: Other: Advisory role, Research Funding. Pemmaraju:Blueprint Medicines: Honoraria; Roche Diagnostics: Honoraria; Incyte Corporation: Honoraria; Novartis: Honoraria, Research Funding; DAVA Oncology: Honoraria; Celgene: Honoraria; Samus Therapeutics: Research Funding; Daiichi Sankyo: Research Funding; AbbVie: Honoraria, Research Funding; Plexxikon: Research Funding; Stemline Therapeutics: Honoraria, Research Funding; LFB Biotechnologies: Honoraria; Cellectis: Research Funding; MustangBio: Honoraria; Pacylex Pharmaceuticals: Consultancy; SagerStrong Foundation: Other: Grant Support; Affymetrix: Other: Grant Support, Research Funding. Kadia:Pulmotec: Research Funding; Astra Zeneca: Research Funding; Cellenkos: Research Funding; BMS: Honoraria, Research Funding; Novartis: Honoraria; Pfizer: Honoraria, Research Funding; Abbvie: Honoraria, Research Funding; Ascentage: Research Funding; Astellas: Research Funding; Amgen: Research Funding; Celgene: Research Funding; JAZZ: Honoraria, Research Funding; Genentech: Honoraria, Research Funding; Cyclacel: Research Funding; Incyte: Research Funding. Ravandi:AstraZeneca: Consultancy, Honoraria; Macrogenics: Research Funding; Xencor: Consultancy, Honoraria, Research Funding; Jazz Pharmaceuticals: Consultancy, Honoraria, Research Funding; Abbvie: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria; BMS: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; Astellas: Consultancy, Honoraria, Research Funding; Orsenix: Consultancy, Honoraria, Research Funding. Daver:Trovagene: Research Funding; Fate Therapeutics: Research Funding; ImmunoGen: Research Funding; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees; Jazz: Consultancy, Membership on an entity's Board of Directors or advisory committees; Trillium: Consultancy, Membership on an entity's Board of Directors or advisory committees; Syndax: Consultancy, Membership on an entity's Board of Directors or advisory committees; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; KITE: Consultancy, Membership on an entity's Board of Directors or advisory committees; Agios: Consultancy, Membership on an entity's Board of Directors or advisory committees; Daiichi Sankyo: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bristol-Myers Squibb: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Karyopharm: Research Funding; Servier: Research Funding; Genentech: Research Funding; AbbVie: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Astellas: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novimmune: Research Funding; Gilead: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding. Kantarjian:Janssen: Honoraria; Oxford Biomedical: Honoraria; BMS: Research Funding; Ascentage: Research Funding; Amgen: Honoraria, Research Funding; Abbvie: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Jazz: Research Funding; Delta Fly: Honoraria; BioAscend: Honoraria; Aptitute Health: Honoraria; Adaptive biotechnologies: Honoraria; Actinium: Honoraria, Membership on an entity's Board of Directors or advisory committees; Sanofi: Research Funding; Pfizer: Honoraria, Research Funding; Immunogen: Research Funding; Daiichi-Sankyo: Honoraria, Research Funding. Garcia-Manero:Bristol-Myers Squibb: Consultancy, Research Funding; AbbVie: Honoraria, Research Funding; Helsinn Therapeutics: Consultancy, Honoraria, Research Funding; Acceleron Pharmaceuticals: Consultancy, Honoraria; Amphivena Therapeutics: Research Funding; Novartis: Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Jazz Pharmaceuticals: Consultancy; Merck: Research Funding; Genentech: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Onconova: Research Funding; H3 Biomedicine: Research Funding; Astex Pharmaceuticals: Consultancy, Honoraria, Research Funding. OffLabel Disclosure: Venetoclax for CMML

At higher order levels chromatin fibers in interphase nuclei are organized into loop domains. Gene regulatory elements (promoters and enhancers) are often located near the sites of loop attachments. Therefore, loop domains play a key role in regulation of cell transcriptional activity. We investigated the kinetics of DNA loop exit during single cell gel electrophoresis (the comet assay) of nucleoids obtained from two cell types that differ in their synthetic activity – human lymphocytes and lymphoblasts. Lymphocyte activation and transformation into lymphoblasts (blast transformation) was performed with interleukin 2. The results obtained suggest that a rearrangement of the loops occurs after lymphocyte activation. After blast transformation we observed an increase of the amount of loop domains on the surface of nucleoids against a decrease of the inner loop fraction. Therefore, the comet assay can be used for detection of large-scale changes in the cell nucleus that follow changes in cell functional state.

The purpose of this study was to monitor abundance and activation of local CD8β-expressing T-cell populations during Eimeria tenella infections of naïve chickens and chickens immune by previous infections. Chickens were infected with E. tenella up to three times. Caecal T-cell receptor (TCR) γ/δ-CD8β+ cells (cytotoxic T lymphocytes; CTL) and TCRγ/δ+CD8β+ cells were characterized with respect to activation markers (blast transformation, CD25 and cell surface CD107a). Cells were also induced to degranulate in vitro as a measure of activation potential. Major findings included a prominent long-lasting, up to 6 weeks, increase in the proportion of CTL among caecal CD45+ cells in the later stages after primary E. tenella infection. These CTL also showed clear signs of activation, that is blast transformation and increased in vitro induced degranulation. At second and third E. tenella infection, chickens showed strong protective immunity but discrete signs of cellular activation were observed, for example increased in vitro induced degranulation of CTL. Thus, primary E. tenella infection induced clear recruitment and activation of local CTL. Upon subsequent infections of strongly immune chickens cellular changes were less prominent, possibly due to lower overall numbers of cells being activated because of the severe restriction of parasite replication.

The evolution to blast phase is a frequently unpredictable and almost invariably fatal event in the course of myeloproliferative neoplasms. The molecular mechanisms underlying blast transformation have not been elucidated and the specific genetic and epigenetic events governing leukemogenesis remain unclear. The result of the long-lasting dynamics, passing through progressive genetic steps, is the emergence of one or more clones often characterized by complex genetics, either at conventional karyotyping or at modern high-throughput sequencing analyses, with all clinical and prognostic correlates. The current therapeutic approaches are largely inadequate and incapable of modifying the inherent unfavorable outcome. In this perspective, the application of targeted strategies should aim to prevent the occurrence of leukemic evolution. At transformation, the crucial target of treatment should be the allocation to allogeneic transplant for eligible patients. With this in mind, novel combination treatments may provide useful bridging strategies, beyond potentially improving outcomes for patients who are not candidates for intensive approaches.

Among 274 patients with chronic myelomonocytic leukemia (CMML) and followed for a median of 17.1 months, blast transformation (BT) occurred in 36 (13%). On multivariable analysis, risk factors for BT were presence of circulating blasts (HR 5.7; 95% CI 2.8–11.9) and female gender (HR 2.6; 95% CI 1.3–5.1); the results remained unchanged when analysis was restricted to CMML-1. ASXL1/SRSF2/SF3B1/U2AF1/SETBP1 mutational frequencies were not significantly different between time of CMML diagnosis and BT. Median survival post-BT was 4.7 months (5-year survival 6%) and better with allogeneic stem cell transplant (SCT) (14.3 months vs. 4.3 months for chemotherapy vs. 0.9 months for supportive care; P = 0.03). Neither karyotype nor mutational status was independently associated with risk of BT or post-BT survival. We conclude that female patients with CMML and those with circulating blasts are at a higher risk of BT. Post-BT survival is dismal and our observations suggest consideration of allogeneic SCT prior to BT.Am. J. Hematol. 90:411–416, 2015. © 2015 Wiley Periodicals, Inc.

Lymphocytes respond to mitogens that stimulate proliferation by increasing theirs metabolic activity. In this study we investigate L-Glutamine and L-Leucine uptake as markers of cell response to Concavalina A (ConcaA) stimulation, using a high-resolution flow technique. We found that lymphocytes induced to blast transformation enhanced rate and efficiency of amino acid uptake during cell proliferation. Considering that increases in transport is the first quantifiable response of cells during malignant transformation, amino acid uptake could also be useful as an early marker of malignancy.

Лимфоцити од болни со тиротоксикоза инкубирани со тироиден целуларен Аг покажуваат бластна трансформација во висок степен.Резултатите укажуваат дека кај тиреотоксикоза постои претходна сензибилизација на лимфоцитите со тироиден антиген што укажува на пореметување на целуларниот имунитет кај овие болни., It has been examined the possibility of thyroid tissue as to how as a same antigen influence on blast - transformation, eventually on previously sensitised lymphocytes at thyrotoxicosis patients.The lymphocytes have been separated (taken) from the vein blood by the method of centrifugation with ficoll. Upon checking the lymphocytes are incubated in the medium in addition to homogenised thyroid tissue.It has been demonstrated the relation between the unchanged and blast transformed lymphocytes after 24 h incubation of the lymphocytes in the medium with the thyroid antigen.

Lymphocyte proliferation in response to antigenic or mitogenic stimulation is a readily quantifiable phenomenon useful for testing immunomodulatory (i.e., immunosuppressive or immunostimulatory) chemical compounds and biologics. One of the earliest steps during mitogenesis is cell enlargement or blastogenic transformation, whereupon the cell volume increases before division. It is usually detectable in the first several hours of T-lymphocyte stimulation. Here, we describe a rapid method to quantify blastogenesis in T lymphocytes isolated from mouse spleens and human peripheral blood mononuclear cells (PBMCs) using an automated cell counter. Various commonly used proliferation assays for the most part are laborious and only reflect the overall population effect rather than individual cellular effects within a population. In contrast, the presented automated cell counter assay provides rapid, direct, and precise measurements of cell diameters that can be used for assessing the effectiveness of various mitogens and immunomodulatory drugs in vitro.

Eleven thrombocytopenic members of a large family with sex-linked hereditary thrombocyto-penia were studied. Routine immunological investigation revealed little evidence of immune deficiency, but abnormal results were obtained in studies of the in vitro blast transformation response of the patients lymphocytes. Impaired responses to microbial antigens were observed in all patients thus studied, whereas a decreased response to mitogens was observed in only one patient. Of the non-thrombocytopenic family members, approximately half showed subnormal responses to one microbial extract. These results, taken together with the increased incidence of infections in the thrombocytopenic patients, indicate the necessity for immunological scrutiny of patients with hereditary thrombocytopenia, in particular when splenectomy is considered.

Zusammenfassung: Es wird ein 20jahriger Patient aus Mozambique mit einer nordamerikanischen Blastomykose n. Bl. vorgestellt. Die Verabreichung von Ketoconazol fuhrtezu einer Besserung des klinischen Bildes, jedoch nicht zur Abheilung. Als Mittel der Wahl erwies sich nach chirugischer Exstirpation beteiligter Rippenanteile eine Kombinationstherapie von Amphotericn B i.v. + 5-Flucytosin. Immunologische Parameter wurden vor und am Ende der Therapie gepruft: Der Intrakutantest mit Varidase, PPD, Tkichosan, Histoplasmin und Blastomycin war positiv. Dysreguiationen unspezifiscber Abwehrmechanismen und zellvermittelter Immunreaktionen konnten nachgewiesen werden. Die Leukozytenadharenz war sehr stark herabgesetzt, die Zahl E+ -T-Zellen erniedrigt. Im LTT fehlte die T-Zellmitogenreaktivitat vollstandig; monozytare Suppressionseffekte”kurzlebige Suppressorellen“konnten nicht registriert werden. Im Leukozytenmigrationstest war mit Trichophytin, Candidin und Histoplasmin eine Hemmung der Migration gegen diese Antigene nachweisbar LMT 1, 2. Mit Blastomycin war keine Sensibilisierung erkennbar. Nach der Behandlung regiderten wir weiterhin eine Abnahme der Phagozytoserate weit unter den Normbereich beigegenlaunfiger Tendenz des Phagozytoseindex. - Im LTT waren jetzt T-Zellmitogeneffekte erkennbar, der Normbereich de nicht erreicht. PPD und Candida-Antigen fuhrten zu einer erheblichen Stimulierung der Blasttransformation; die Histoplasmin-Stimulierbarkeit nahm unter der Therapie ab. Monozytare Suppressoreffekte konnten jetzt verstarkt nachgewiesen werden SZI 4.0. Die immunologischen Befunde werden als kombinierter T-Zell- und Monozytendefekt gewertet und diskutiert. Summary: A 20-year-old patient from Mozambique with a North American Blastomycosis n. BL was admitted for treatment. The administration of ketoconazole brought a clinical improvement, but did not heal the patient. Combined therapy with amphotericin B+5-flucytosine following surgical exstirpation proved successful. Immunological parameters were tested before and after therapy: The intracutaneous tests with varidase, PPD, tricbosan, histoplasmin and blastomycin were positive. Dysregulation of nonspecific and specific immune responses was detected. The leucocyte adherence LA was substantially, and the number of T-cells slightly, reduced. In the LTT the T cell mitogen reactivity was absent. No monocytic suppression effects (shortlived suppressor cells) could be detected. The leucocytes migration test LMT with trichophytin, candidin and histoplasmin revealed a specifrc sensitization LMT l, 2. This sensitization could not be detected with blastomycin. After therapy we also observed a reduction in the phagocytosis rate far below the normal range, the phagocytosis index showing the opposite tendency. T-cell mitogenic effets now also appeared in the LTT, but the normal range was not reached. PPD and Candida-antigen continued to induce substantial stimulation of blast transformation. Monocytic suppressor effects were more pronounced SZI 4.0. - The immunological results were discussed as a combined T cell and monocyte defect.