CD4 Cells In Patients Research Articles

Deteksi HIV / AIDs Dengan Pemeriksaan Viral Load Dan Pemeriksaan Penghitungan CD4 Cell

The incidence of cases of HIV / AIDS is still increasing, including in South Kalimantan, especially Banjarmasin, which has an incidence of cases of HIV / AIDS in 2018 totaling 181 people. The HIV virus attacks white blood cells (T-helper lymphocytes "CD4 +) and results in decreased human immunity. The viral load examination is used to measure the amount of HIV virus in the blood. Examination of CD4 cell counts are used to assess the level of immunity of people with HIV / AIDS. The purpose of this study was to determine the relationship between the results of the viral load examination with the results of the examination of the number of CD4 cells in patients with HIV / AIDS. This research can provide scientific information as a reference to increase knowledge about HIV / AIDS and decrease the level of immunity of people with HIV / AIDS. This research also provides insight into how to find out initial information about the body's immunity against new sufferers. Sampling was done by purposive sampling with inclusion criteria. Samples are new outpatients, aged 20-50 years and have received VCT (Voluntary counseling and testing). Sample is an HIV / AIDS positive HIV patient by conducting an HIV rapid test. Determination of the sample was also seen from the exclusion criteria of HIV / AIDS sufferers who had received ARV drug therapy. This research was an observational analytic study with a cross-sectional approach using quantitative analysis methods. Viral load examination uses the real time reverse transcriptase method - Polymerase Chain Reaction (RT-PCR). Examination of CD4 cell numbers using the flowcytometry method with the Alere PIMA tool. Data were analyzed by Rank Spearman correlation test. The results of the Viral load in patients with HIV / AIDS in the adult age group showed 9,000 copies / ml and at the end 960,000 copies / ml. The results of the viral load test in adult patients at the end are higher than at the beginning. Examination Results of the number of CD4 cells in patients with HIV / AIDS in the adult age group showed 586 cells / ul and at the end of 11 cells / ul. Examination Results of CD4 cell counts for adult sufferers show lower end than at baseline. Indications of the results of the examination showed the higher the amount of viral load, the number of cells CD4 will be lower than patients with HIV / AIDS. The implication of this research is a recommendation of viral load examination for detection new patient. The conclusion of this study is that there is a significant relationship between the results of the examination of viral load and the results of the examination of the number of CD4 cells in patients with HIV / AIDS.

Skewing of Sodium Antimony Gluconate mediated therapy for a predominant Th1 during visceral leishmaniasis on triggering CD2 epitope to circumvent liver infection

Visceral leishmaniasis (VL) is a macrophage associated disorder which is linked with a profound decrease in the immunotherapeutic potential of the infected subjects leading to a marked reduction in the CD4 linked Th1 protective immune response. It greatly affects the liver leading to abnormal levels of serum glutamic pyruvic transaminase and serum glutamic oxaloacetic transaminase. Also the patients suffering from VL have been reported to be coinfected with Hepatitis C during some circumstances. Simultaneously the patients in Bihar are showing unresponsiveness towards sodium antimony gluconate which is still a first line of drug in many countries around the world against VL. The T cells were isolated from patients and were subjected to fluroscence associated cell sorting analysis and cytokine analysis. We have reported down regulation of CD2 co receptor on the surface of CD4 cells in patients suffering from VL. Stimulation of CD2 epitope with anti CD2 antibody has led to a remarkable increase in the protein kinase C alpha mediated phosphorylation on CD2 co receptor on CD4 T cells, induction of interferon (IFN)-γ led Th1 dominated immune response, a substantial increase in the lymphoblast population and this response remained Th1 dominated even in the presence of Th2 predominant conditions signified with recombinant interleukin-4. We have shown that biological immunomodulators such as IFN-γ can provide a missing signal and enhance the activity of antimonials in the treatment of VL and cutaneous leishmaniasis.

136P - Evaluation of a radiomic signature of CD8 cells in patients treated with immunotherapy-radiotherapy in three clinical trials

BackgroundSeveral studies have suggested that combining radiotherapy (RT) to immunotherapy (IO) may be synergistic but many questions are still pending regarding the radiation modalities to optimize this combination, such as the choice of the lesion to irradiate. Radiomics consists in the analysis of quantitative data extracted from standard medical imaging to generate imaging biomarkers. A previous study published in The Lancet Oncology has shown that a radiomic signature could predict the CD8 cells infiltration, which is associated with the activity of anti-PD-1/PD-L1. We aimed to assess whether this biomarker could help to guide IO-RT combinations. MethodsPatients from three clinical studies of IO-RT combinations with advanced solid tumors in two institutions were screened. Patients with available baseline (E0) and first evaluation (E1) CTs were included. Immunotherapy consisted in 4 different drugs. Hypofractionated conformal RT or stereotactic RT of one tumor lesion was delivered after the start of IO for most of the patients. The irradiated lesion and a sample of non-irradiated lesions were delineated from E0 and E1 CTs. Radiomics features were extracted and the published radiomic signature was applied to estimate the CD8 cells. Results84 patients were included. 244 tumor lesions were delineated on the E0 CT, including the 84 lesions which were selected for irradiation. Median time between IO and RT start was 21 days (IQR: 9-24), and 2.4mo between E0 and E1 (IQR: 1.3 - 3). 80 irradiated lesions and 152 non irradiated lesions remained at E1. At baseline, the volume and the radiomic score of TIL (RS) were not different between the two groups (irradiation or no) (p=0.94 and 0.50). While the mean volume of the analyzed lesions was not different from E1 to E0 (p=0.15), irradiated lesions were significantly smaller at E1 (p=0.03). A high RS in the irradiated lesion at E1 (compared to the median value) was associated with PFS (HR=0.57, IC95%: 0.345-0.95, p=0.031) irrespective of the volume in multivariate analysis but was not significantly associated with OS. ConclusionsRadiomic score of the irradiated lesion was associated with PFS. Such biomarker may help to guide the selection of the lesion to irradiate in IO-RT combinations. Legal entity responsible for the studyGustave Roussy Cancer Campus. FundingFondation pour la Recherche Médicale, SIRIC-SOCRATE 2.0, Fondation ARC, Amazon. DisclosureR. Sun: Travel / Accommodation / Expenses: AstraZeneca. N.L. Sundahl: Travel / Accommodation / Expenses: Merck Sharpe & Dohme; Travel / Accommodation / Expenses: Astellas; Travel / Accommodation / Expenses: Bayer; Travel / Accommodation / Expenses: Bristol-Myers Squibb. P. Ost: Research grant / Funding (institution): Merck Sharpe & Dohme; Research grant / Funding (institution): Astellas; Research grant / Funding (institution): Janssen; Travel / Accommodation / Expenses: Ipsen; Honoraria (self), Travel / Accommodation / Expenses: Ferring Pharmaceuticals; Honoraria (self): Bayer. C. Massard: Advisory / Consultancy: Amgen; Advisory / Consultancy: AstraZeneca; Advisory / Consultancy: Bayer; Advisory / Consultancy: Celgene; Advisory / Consultancy: Genentech; Advisory / Consultancy: Ipsen; Advisory / Consultancy: Jansen; Advisory / Consultancy: Lilly; Advisory / Consultancy: Novartis; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Roche; Advisory / Consultancy: Sanofi; Advisory / Consultancy: Orion. E. Deutsch: Advisory / Consultancy, Research grant / Funding (institution): Roche Genentech; Advisory / Consultancy, Research grant / Funding (institution): Merck Serono; Research grant / Funding (institution): AstraZeneca; Research grant / Funding (institution): BMS; Research grant / Funding (institution): MSD. All other authors have declared no conflicts of interest.

The clinical significance of the changes of Th17 cells and cytokines IL-17 and IL-6 in hepatocellular carcinoma patients

Objective To investigate the change of the percentage of Th17 cells and cytokine among hepatocellular carcinoma (HCC) tissues and para-carcinoma tissue and the liver tissue of hepatic hemangioma, and to explore its clinical significance. Methods To choose 26 of hepatocellular carcinoma patients between November 2014 and December 2016 and also include 11 cases of hepatic hemangioma as control group. The percentage of Th17cells in HCC tissue and para-carcinoma tissue and the liver tissue of hepatic hemangioma was evaluated by flow cytometric; The levels of IL-6 and IL-17 in HCC tissues, para-carcinoma tissue and the liver tissue of hepatic hemangioma were detected by RT-PCR. Results The ratio of Th17 cells to CD4 cells in peripheral blood of HCC was significantly higher than that in control group (4.07%±0.68% vs. 1.39%±0.41%), P<0.05. The ratio of Th17 cells to CD4 cells in patients with hepatocellular carcinoma was significantly higher than that in paracancerous tissues and liver tissues of hepatic hemangioma (4.76%±0.75% vs. 2.30%±0.26% vs. 0.77%±0.31%, P<0.05). Multivariate linear stepwise regression analysis of the changes of Th17 cells in patients with HCC and their correlation with clinical parameters showed that Th17 lymphocytes in peripheral blood of patients with HCC were closely related to TNM staging(YTh17=2.647+ 0.687TNM. It’s indicated that the level of IL-6 and IL-17 in liver cancer tissues was significantly higher than that of the liver hemangioma in the control group through ELISA (P<0.05). The proportion of peripheral blood Th17 cells to CD4 cells is different in different TNM stages(stage I<stage II<stage III~IV, the difference was statistically significant. RT-PCR analysis showed that the expression of IL-17, IL-6mRNA in hepatocellular carcinoma was significantly higher than that in adjacent tissues and hepatic hemangioma liver tissue (all P<0.05). Conclusion The increased proportion of Th17 cells and the increased levels of cytokines IL-6 and IL-17 may be closely correlated with occurrence and development of hepatocellular carcinoma. It can be an important target of anti-tumor therapy. Key words: Liver neoplasms; Carcinoma, hepatocellular; Th17 cells; Interleukin-6; Interleukin-17

Impact of HBV and HCV coinfection on CD4 cells among HIV-infected patients: a longitudinal retrospective study

The impact of hepatitis B virus (HBV) and hepatitis C virus (HCV) coinfection on CD4 cells in patients with human immunodeficiency virus (HIV) is unclear. We aimed to examine the impact of HBV and HCV coinfection on CD4 cell count and CD4/CD8 ratio in adults with HIV. We conducted a longitudinal retrospective study in Brazil between January 1, 2002, and June 30, 2016, including 205 patients with HIV monoinfection, 37 with HIV-HBV coinfection, 35 with HIV-HCV coinfection, and 62 with HIV-HCV (48 HCV genotype 1 and 14 HCV genotype 3). Median duration of follow-up was 2,327 (interquartile range: 1,159-3,319) days. An increased CD4 cell count and CD4/CD8 ratio over time was observed in all groups receiving combined antiretroviral therapy (cART). Patients with HIV-HBV or HIV-HCV coinfection and those with HIV monoinfection, showed comparable CD4 cell counts and CD4/CD8 ratios during pre-ART. There was also no statistically significant difference in CD4/CD8 ratio between HIV-HBV or HIV-HCV coinfection groups and the HIV monoinfection group during follow-up on cART. However, CD4 cell counts were significantly lower in HIV-HCV patients than in HIV monoinfection patients during follow-up on cART. HIV patients with HCV genotype 3 coinfection showed significantly lower CD4/CD8 ratio during follow-up on cART than those coinfected with HCV genotype 1 coinfection. No statistically significant effect of coinfection was observed on the efficacy of cART. HIV-infected patients are more likely to show better immunological responses to cART when they are not coinfected with HCV.

A radiomics approach to assess tumour-infiltrating CD8 cells and response to anti-PD-1 or anti-PD-L1 immunotherapy: an imaging biomarker, retrospective multicohort study.

Because responses of patients with cancer to immunotherapy can vary in success, innovative predictors of response to treatment are urgently needed to improve treatment outcomes. We aimed to develop and independently validate a radiomics-based biomarker of tumour-infiltrating CD8 cells in patients included in phase 1 trials of anti-programmed cell death protein (PD)-1 or anti-programmed cell death ligand 1 (PD-L1) monotherapy. We also aimed to evaluate the association between the biomarker, and tumour immune phenotype and clinical outcomes of these patients. In this retrospective multicohort study, we used four independent cohorts of patients with advanced solid tumours to develop and validate a radiomic signature predictive of immunotherapy response by combining contrast-enhanced CT images and RNA-seq genomic data from tumour biopsies to assess CD8 cell tumour infiltration. To develop the radiomic signature of CD8 cells, we used the CT images and RNA sequencing data of 135 patients with advanced solid malignant tumours who had been enrolled into the MOSCATO trial between May 1, 2012, and March 31, 2016, in France (training set). The genomic data, which are based on the CD8B gene, were used to estimate the abundance of CD8 cells in the samples and data were then aligned with the images to generate the radiomic signatures. The concordance of the radiomic signature (primary endpoint) was validated in a Cancer Genome Atlas [TGCA] database dataset including 119 patients who had available baseline preoperative imaging data and corresponding transcriptomic data on June 30, 2017. From 84 input variables used for the machine-learning method (78 radiomic features, five location variables, and one technical variable), a radiomics-based predictor of the CD8 cell expression signature was built by use of machine learning (elastic-net regularised regression method). Two other independent cohorts of patients with advanced solid tumours were used to evaluate this predictor. The immune phenotype internal cohort (n=100), were randomly selected from the Gustave Roussy Cancer Campus database of patient medical records based on previously described, extreme tumour-immune phenotypes: immune-inflamed (with dense CD8 cell infiltration) or immune-desert (with low CD8 cell infiltration), irrespective of treatment delivered; these data were used to analyse the correlation of the immune phenotype with this biomarker. Finally, the immunotherapy-treated dataset (n=137) of patients recruited from Dec 1, 2011, to Jan 31, 2014, at the Gustave Roussy Cancer Campus, who had been treated with anti-PD-1 and anti-PD-L1 monotherapy in phase 1 trials, was used to assess the predictive value of this biomarker in terms of clinical outcome. We developed a radiomic signature for CD8 cells that included eight variables, which was validated with the gene expression signature of CD8 cells in the TCGA dataset (area under the curve [AUC]=0·67; 95% CI 0·57-0·77; p=0·0019). In the cohort with assumed immune phenotypes, the signature was also able to discriminate inflamed tumours from immune-desert tumours (0·76; 0·66-0·86; p<0·0001). In patients treated with anti-PD-1 and PD-L1, a high baseline radiomic score (relative to the median) was associated with a higher proportion of patients who achieved an objective response at 3 months (vs those with progressive disease or stable disease; p=0·049) and a higher proportion of patients who had an objective response (vs those with progressive disease or stable disease; p=0·025) or stable disease (vs those with progressive disease; p=0·013) at 6 months. A high baseline radiomic score was also associated with improved overall survival in univariate (median overall survival 24·3 months in the high radiomic score group, 95% CI 18·63-42·1; vs 11·5 months in the low radiomic score group, 7·98-15·6; hazard ratio 0·58, 95% CI 0·39-0·87; p=0·0081) and multivariate analyses (0·52, 0·35-0·79; p=0·0022). The radiomic signature of CD8 cells was validated in three independent cohorts. This imaging predictor provided a promising way to predict the immune phenotype of tumours and to infer clinical outcomes for patients with cancer who had been treated with anti-PD-1 and PD-L1. Our imaging biomarker could be useful in estimating CD8 cell count and predicting clinical outcomes of patients treated with immunotherapy, when validated by further prospective randomised trials. Fondation pour la Recherche Médicale, and SIRIC-SOCRATE 2.0, French Society of Radiation Oncology.

Phenotypes and HIV-1-specific T cell responses of KIR3DL1 positive CD8 cells in patients with early HIV-1 infection

Objective To investigate the phenotypes and the HIV-1-specific T cell responses of KIR3DL1 positive CD8 cells in patients with early HIV-1 infection. Methods Fifty-six HIV-1 antibody negative individuals and thirty-two patients with early HIV-1 infection were enrolled in the study. Fluorescence-activated cell sorting (FACS) was performed to detect the phenotypes of KIR3DL1 receptor expressed on the surface of CD8 cells. The levels of IFN-γ were measured by intracellular cytokine staining assay after the PBMCs were stimulated with an HIV-1 Gag peptide pool. Results The percentages of KIR3DL1+ CD8 T cells in HIV-1 negative individuals and patients with early HIV-1 infection were 1.45% (0.12%-8.4%) and 0.82% (0.14%-6.14%), respectively, and there was no significant difference between them. The percentages of KIR3DL1+ CD8 Temra cells in HIV-1 negative individuals and patients with early HIV-1 infection were (4.55±3.84)% and (6.71±8.50)%, respectively, which were significantly higher than the percentages of KIR3DL1+ CD8 Tem cells, which were (0.50±0.59)% and (1.18±1.39)%, respectively (all P<0.01). Moreover, the percentages of KIR3DL1+ CD8 Tem cells in patients with early HIV-1 infection were higher than those in HIV-1 negative individuals (P=0.001 2). The percentage of KIR3DL1+ CD8 Temra cells was positively correlated with the HIV-1 viral load in patients with early HIV-1 infection (rs=0.576, P=0.000 9). The percentages of KIR3DL1+ CD8 Temra cells in HIV-1 patients, whose viral loads were larger than 4.0log, were much higher than those in HIV-1 patients with viral loads less than 4.0 log (P=0.002). Additionally, the levels of IFN-γ secreted by KIR3DL1 positive CD8 cells were much lesser than those secreted by KIR3DL1 negative CD8 cells (P<0.000 1). Conclusion The receptor of KIR3DL1 was mainly expressed on CD8 Temra cells in both HIV-1 negative subjects and patients with early HIV-1 infection. High HIV-1 viremia was associated with the high percentage of KIR3DL1+ CD8 Temra cells. The KIR3DL1 positive CD8 cells induced lower HIV-1-specific T cell responses. Key words: Early HIV-1 infection; KIR3DL1 receptor; CD8 cells

Residual Plasma Viraemia and Infectious HIV-1 Recovery from Resting Memory CD4 Cells in Patients on Antiretroviral Therapy: Results from ACTG A5173

In HIV-1-infected patients receiving antiretroviral therapy (ART), the relationship between residual viraemia and ex vivo recovery of infectious virus from latently infected CD4 cells is uncertain. We measured residual viraemia (HIV-1 RNA copies/ml) by single-copy assay (SCA) and the latent reservoir by infectious virus recovery from resting memory CD4 cells (infectious units per million cells [IUPM]) in patients who initiated ART. We assessed immune activation by measuring CD38 expression on T-cells. Ten patients who initiated ART and maintained a plasma HIV-1 RNA level < 200 copies/ml had residual viraemia and IUPM measured every 24 weeks. Five of 10 patients had longitudinal IUPM measured at weeks 24-96; the remainder had IUPM measured 1-3 times over 24-72 weeks. Analyses of 29 paired measurements revealed a positive association between level of residual viraemia and IUPM (0.56 higher log10 HIV-1 RNA copies/ml per 1 log10 higher IUPM; P = 0.005). Residual viraemia level was positively associated with CD38 density and percentage on CD8+ T-cells in concurrent samples and with pre-ART HIV-1 RNA levels. In patients with HIV-1 RNA levels < 200 copies/ml 24-96 weeks after initiating ART, the level of viraemia is positively associated with infectious virus recovery from resting memory CD4 cells. Whether this association persists after longer-term suppressive ART needs to be determined. If additional studies show that residual viraemia measured by SCA reflects the size of the latent reservoir in patients who have had virological suppression for longer periods of time, this could facilitate testing of potentially curative strategies to reduce this important reservoir.

Increased Frequencies of Myelin Oligodendrocyte Glycoprotein/MHC Class II-Binding CD4 Cells in Patients with Multiple Sclerosis

Multiple sclerosis (MS) is an autoimmune disease characterized by infiltration of pathogenic immune cells in the CNS resulting in destruction of the myelin sheath and surrounding axons. We and others have previously measured the frequency of human myelin-reactive T cells in peripheral blood. Using T cell cloning techniques, a modest increase in the frequency of myelin-reactive T cells in patients as compared with control subjects was observed. In this study, we investigated whether myelin oligodendrocyte glycoprotein (MOG)-specific T cells could be detected and their frequency was measured using DRB1*0401/MOG(97-109(107E-S)) tetramers in MS subjects and healthy controls expressing HLA class II DRB1*0401. We defined the optimal culture conditions for expansion of MOG-reactive T cells upon MOG peptide stimulation of PMBCs. MOG(97-109)-reactive CD4(+) T cells, isolated with DRB1*0401/MOG(97-109) tetramers, and after a short-term culture of PMBCs with MOG(97-109) peptides, were detected more frequently from patients with MS as compared with healthy controls. T cell clones from single cell cloning of DRB1*0401/MOG(97-109(107E-S)) tetramer(+) cells confirmed that these T cell clones were responsive to both the native and the substituted MOG peptide. These data indicate that autoantigen-specific T cells can be detected and enumerated from the blood of subjects using class II tetramers, and the frequency of MOG(97-109)-reactive T cells is greater in patients with MS as compared with healthy controls.

CD8 Cells of Patients with Diffuse Cutaneous Leishmaniasis Display Functional Exhaustion: The Latter Is Reversed, In Vitro, by TLR2 Agonists

Leishmania mexicana (Lm) causes localized (LCL) and diffuse (DCL) cutaneous leishmaniasis. DCL patients have a poor cellular immune response leading to chronicity. It has been proposed that CD8 T lymphocytes (CD8) play a crucial role in infection clearance, although the role of CD8 cytotoxicity in disease control has not been elucidated. Lesions of DCL patients have been shown to harbor low numbers of CD8, as compared to patients with LCL, and leishmanicidal treatment restores CD8 numbers. The marked response of CD8 towards Leishmania parasites led us to analyze possible functional differences between CD8 from patients with LCL and DCL. We compared IFNγ production, antigen-specific proliferation, and cytotoxicity of CD8 purified from PBMC against autologous macrophages (MO) infected with Leishmania mexicana (MOi). Additionally, we analyzed tissue biopsies from both groups of patients for evidence of cytotoxicity associated with apoptotic cells in the lesions. We found that CD8 cell of DCL patients exhibited low cytotoxicity, low antigen-specific proliferation and low IFNγ production when stimulated with MOi, as compared to LCL patients. Additionally, DCL patients had significantly less TUNEL+ cells in their lesions. These characteristics are similar to cellular “exhaustion” described in chronic infections. We intended to restore the functional capacity of CD8 cells of DCL patients by preincubating them with TLR2 agonists: Lm lipophosphoglycan (LPG) or Pam3Cys. Cytotoxicity against MOi, antigen-specific proliferation and IFNγ production were restored with both stimuli, whereas PD-1 (a molecule associated with cellular exhaustion) expression, was reduced. Our work suggests that CD8 response is associated with control of Lm infection in LCL patients and that chronic infection in DCL patients leads to a state of CD8 functional exhaustion, which could facilitate disease spread. This is the first report that shows the presence of functionally exhausted CD8 T lymphocytes in DCL patients and, additionally, that pre-stimulation with TLR2 ligands can restore the effector mechanisms of CD8 T lymphocytes from DCL patients against Leishmania mexicana-infected macrophages.

No Evidence for Decay of the Latent Reservoir in HIV‐1–Infected Patients Receiving Intensive Enfuvirtide‐Containing Antiretroviral Therapy

Human immunodeficiency virus type 1 (HIV-1) persists in a latent reservoir of infected resting memory CD4 cells in patients receiving antiretroviral therapy. We assessed whether multitarget therapy with enfuvirtide, 2 reverse-transcriptase inhibitors, and a ritonavir-boosted protease inhibitor leads to decay of this reservoir. Nineteen treatment-naive patients initiated this regimen; 9 experienced virologic suppression and continued enfuvirtide-containing therapy for at least 48 weeks. In enfuvirtide-treated patients with virological suppression, there was no decay of the latent reservoir (95% confidence interval for half-life, 11 months to infinity). The stability of the latent reservoir despite intensive therapy suggests that new strategies are needed to eradicate HIV-1 from this reservoir. (ClinicalTrials.gov identifier: NCT00051831 .).

Defect of regulatory T cells in patients with Omenn syndrome

Omenn syndrome (OS) is an autosomal-recessive disorder characterized by severe immunodeficiency and T-cell-mediated autoimmunity. The disease is caused by hypomorphic mutations in recombination-activating genes that hamper the process of Variable (V) Diversity (D) Joining (J) recombination, leading to the generation of autoreactive T cells. We have previously shown that in OS the expression of autoimmune regulator, a key factor governing central tolerance, is markedly reduced. Here, we have addressed the role of peripheral tolerance in the disease pathogenesis. We have analyzed forkhead box protein P3 (FOXP3) expression in peripheral blood T cells of 4 patients with OS and in lymphoid organs of 8 patients with OS and have tested the suppressive activity of sorted CD4(+) CD25(high) peripheral blood T cells in 2 of these patients. We have observed that CD4(+)CD25(high)T cells isolated ex vivo from patients with OS failed to suppress proliferation of autologous or allogenic CD4(+) responder T cells. Moreover, despite individual variability in the fraction of circulating FOXP3(+) CD4 cells in patients with OS, the immunohistochemical analysis of FOXP3 expression in lymph nodes and thymus of patients with OS demonstrated a severe reduction of this cell subset compared with control tissues. Overall, these results suggest a defect of regulatory T cells in OS leading to a breakdown of peripheral tolerance, which may actively concur to the development of autoimmune manifestations in the disease.

Suppression of Cyclin D 1 (CD1) by on 01910.Na Is Associated with Decreased Survival or Trisomy 8 Myelodysplastic Bone Marrow: A Potential Targetted Therapy for Trisomy 8 MDS.

CD34 positive cells from patients with trisomy 8 myelodysplastic syndrome (MDS) have pronounced expression of early apoptotic markers compared to normal hematopoietic cells. However, trisomy 8 clones persist in patients with bone marrow failure and expand following immunosuppression (Sloand EM et al; Blood 2005; 106(3):841). We have demonstrated up-regulation of c-myc, survivin, and CD1 in CD34 cells of patients with trisomy 8 (Sloand et al; Blood 2007; 109(6):2399). Employing siRNA mediated knockdown of the anti-apoptotic protein survivin, we demonstrated a decrease in trisomy 8 cell growth and postulated that increased Cyclin D1 caused the upregulation of survivin resulting in resistance of these cells to apoptosis.Using fluorescent in situ hybridization (FISH) we showed that the novel styryl sulfone, ON 01910.Na (Vedula MS et al; European Journal of Medicinal Chemistry 2003; 38:811), inhibits cyclin D1 accumulation and is selectively toxic to trisomy 8 cells while promoting maturation of diploid cells. Flow cytometry of cultured cells demonstrated increased proportions of mature CD15 positive myeloid cells and decreased number of immature CD33+ cells or CD34+ blasts (Sloand EM et al; Blood 2007; 110:822).These encouraging in vitro data led to a phase I/II trial of ON 01910.Na in MDS patients with refractory anemia with excess blasts who had IPSS =/> int-2. This study was designed to assess the safety, and activity of escalating doses of ON 01910.Na (800 mg/m2/day × 3 days, 800 mg/m2/day × 5 days, 1500 mg/m2/day × 5 days, 1800 mg/m2/day × 5 days every 2 weeks) in MDS patients. To date five MDS patients have been treated with ON 01910.Na for 4 to 16 weeks in the first two dose cohorts. Two patients had isolated trisomy 8, two had complex cytogenetic abnormalities including trisomy 8 in all aneuploid cells, and one had monosomy 7. Three and five day infusions were well tolerated. Pharmakokinetic analysis showed that the half life of the drug is 1.3 ± 0.5 hours without signs of drug accumulation. Four of five patients demonstrated a rapid and significant decrease in the number of peripheral blasts and aneuploid cells after 4 weeks of therapy (see below), concomitantly with increases in neutrophil and/or platelet counts in four patients. All four patients exhibiting a biological effect of drug treatment had trisomy 8 in their aneuploid clone prior to therapy. One monosomy 7 patient, previously refractory to EPO became responsive to Darbopoietin and another trisomy 8 patient became platelet-transfusion independent.In this early safety trial, ON 01910.Na demonstrates efficacy at early timepoints with respect to improved cytopenias and decreased blast counts. Continued enrollment and long term follow-up will further detail clinical efficacy and impact on the long term prognosis of high risk MDS patients treat with this drug. [Display omitted]

Evidence for the presence of activated CD4 T cells with naive phenotype in the peripheral blood of patients with rheumatoid arthritis.

We have investigated whether T cell activation in rheumatoid arthritis (RA) preferentially engages distinct T cell subpopulations in the peripheral blood (PB) and in the synovial fluid. We found that CD25 expression was enhanced among PB CD4 T cells of RA patients as compared with CD4 cells of patients with reactive arthritis, degenerative joint disease or of healthy controls. Within the CD4 T lymphocytes subset we found that the CD45RO- (naive) cells selectively in RA displayed higher levels of CD25 protein and of interferon-gamma mRNA expression when compared with the respective subset of all other investigated groups. These results show that in the PB of RA, but not in the PB of the other arthropathies or healthy controls, CD45RO-CD4 T lymphocytes exist which display well-defined signs of activation.

Lack of isohemagglutinin production following minor ABO incompatible unrelated HLA mismatched umbilical cord blood transplantation

While immune hemolysis due to donor isohemagglutinin (IH) production often complicates minor ABO incompatible peripheral blood hematopoietic stem cell transplantation (PBSCT), it is not known if this occurs with umbilical cord blood transplantation (UCBT). We compared IH production and hemolysis following minor ABO allogeneic PBSCT and UCBT. We reviewed 24 ABO minor incompatible allogeneic PBSCTs and 14 ABO minor incompatible UCBTs. Patients were evaluated for donor-derived IH by reverse ABO grouping. Evaluation of hemolysis was based on clinical and laboratory findings of anemia associated with increased RBC transfusion need, concomitant with the appearance of donor-derived IH. Of the 24 ABO minor incompatible allogeneic PBSCTs, 15 produced donor-derived IH from 6 to 88 days following transplantation, with seven of 15 patients exhibiting clinically evident hemolysis. There was no significant difference in days to leukocyte engraftment or infused CD34 cells in patients with or without donor-derived IH. None of the 14 patients receiving ABO incompatible UCBTs showed evidence of donor-derived IH following transplantation with a median follow-up of 60 days. We conclude that donor IHs are not produced in patients undergoing minor ABO incompatible UCBTs suggesting fundamental immunologic differences between allogeneic PBSCT and UCBT.

T cell receptor (TCR) BV gene repertoires and clonal expansions of CD4 cells in patients with HIV infections

Despite extensive investigation, the pathogenesis of T cell depletions that characterize AIDS has not been elucidated. To study this process further, we evaluated T cell antigen receptor beta-chain variable gene (TCRBV) repertoires in peripheral blood lymphocytes (PBL) of 23 HIV-infected patients. Expression levels of 28 TCRBV were determined by multiprobe RNase protection assay after polymerase chain reaction (PCR) amplifications. Abnormal expansions (> 2 s.d. from mean normal values) were frequent in HIV CD4, accounting for 26% of total measured TCRBV in this population. The number and magnitude of abnormalities among individuals were inversely proportional to their CD4 counts (P < 0.012 and P < 0.01, respectively). While abnormalities were not randomly distributed among TCRBV subfamilies, no particular genes were expanded or contracted among all patients. Only 14% of CD8 TCRBV were proportionally expanded (P < 0.01 compared with CD4), and there were limited concordances between paired CD8 and CD4 repertoires among individuals. CDR3 length analyses and TCRBV sequencing showed that most CD4 expansions comprised clonal or oligoclonal populations. Thus, T cell responses in HIV patients are characterized by severe TCRBV biases and clonal expansions among CD4 subsets, and these processes are exaggerated with disease progression. The heterogeneity and oligoclonality of the TCRBV expansions are consistent with responses to HIV-encoded or other conventional antigens rather than superantigenic effects. The presence of CD4 clonal proliferations in these patients may be important in the pathogenesis of HIV, and the absence or reduction of many T cell specificities due to oligoclonal expansions may increase susceptibility to opportunistic infections.

BD Biosciences contributions in CD4 counting and immune status for HIV/AIDS.

BD Biosciences is a leader in the use of flow cytometry for determining immune system status and for counting CD4 cells in patients with human immunodeficiency virus (HIV) infection. The company has gained this position through many years of basic research and product development in immunology and cell biology, dye chemistry, immunoassays, instrumentation, and software. Some of the highlights of these developments and their historical perspective are described in this review.

T-helper (Th)1/Th2 imbalance in patients with previously untreated B-cell diffuse large cell lymphoma.

T-helper (Th)1/Th2 imbalance has been observed in a variety of pathological conditions, including malignant diseases. We evaluated the Th1/Th2 balance in peripheral blood Th cells by means of intracellular cytokine analysis in 19 patients with previously untreated B-cell diffuse large cell lymphoma (DLCL) and in 18 patients with B-cell DLCL who had achieved complete remission (CR) after chemotherapy. The mean percentage of Th2 in CD4 cells in patients with DLCL (5.00 +/- 2.20) and that of Th1 in CD4+ cells in patients in CR (32.42 +/- 11.30) were significantly increased in comparison with those in healthy volunteers, respectively (Th1; 23.02 +/- 9.45, Th2; 3.25 +/- 0.90; P<0.01). The mean ratio of Th1/Th2 was significantly lower in patients with DLCL (4.74 +/- 0.52) than in patients in CR (9.31 +/- 1.06; P<0.01) and in healthy volunteers (7.25 +/- 0.65; P<0.01). We conclude that the Th1/Th2 balance was polarized to Th2 in untreated DLCL patients and to Th1 in patients in CR, which suggests that a Th1/Th2 imbalance could play a role in lymphomagenesis and durable remission.

Differential expression profiles of apoptosis-affecting genes in HIV-infected cell lines and patient T cells.

To clarify the molecular mechanisms of HIV-induced apoptosis. The assessment of expression patterns for genes affecting the interrelated cell cycle and apoptosis processes in HIV-1LAI-infected T lymph oblastoid (CEM) cells, as well as CD4 and CD8 cells from HIV-infected individuals and controls. The kinetics of HIV infection in CEM cells were defined by flow cytometry of green fluorescent protein expression from a reporter vector. Apoptosis of CEM cells was measured by propidium iodine staining and flow cytometry. Gene expression levels were determined by a multiprobe RNase protection assay. The infection and apoptosis of CEM cells were associated with enhanced expression of Bax, Bcl-2, Bcl-X(L) and caspase 1 (ICE). There was increased expression of Bcl-2 and caspase 1 and decreased expression of cyclin-dependent kinase inhibitor p21CIP1 in CD4 cells of HIV-infected individuals compared with uninfected controls. The CD8 cells of HIV-infected individuals exhibited increased expression of Bcl-2, Bcl-X(L), Bax and caspase 1 but, in contrast to the CD4 subset, they showed elevated expression of p21CIP1 and p16INK4a compared with controls. The Bax increase in CEM cells appears to be a direct effect associated with a high frequency of infection and apoptosis, because it was not found in the CD4 cells of patients. In contrast, the increase of Bax in the CD8 cells of patients seems to be an indirect effect. Increases in Bcl-2, Bcl-X(L) and caspase 1 in HIV-infected CEM cells may be caused by both direct and indirect mechanisms, because they also occurred in CD4 and CD8 cells of HIV-infected individuals. In addition, the low expression of p21CIP1 in the CD4 subset of HIV-infected individuals could promote apoptosis, whereas the high expression of p21CIP1 and p16INK4a in the CD8 subset may lead to a state of anergy, akin to replicative senescence.

179 Increased number of interferon-gamma producing CD4 and CD8 cells in patients with psoriasis

179 Increased number of interferon-gamma producing CD4 and CD8 cells in patients with psoriasis