CD8 Lymphocyte Subsets Research Articles

Establishment of Reference Values of CD4 and CD8 Lymphocyte Subsets in Healthy Nigerian Adults

A total of 2,570 apparently healthy human immunodeficiency virus-negative adults from the six geopolitical zones in the country were enrolled in our study in 2006. The samples were assayed using the Cyflow technique. Data were analyzed using the Statistical Package for Social Scientists (SPSS). The majority (64%) of the participants had CD4 counts within the range of 501 to 1,000 cells/microl. The reference range for CD4 was 365 to 1,571 cells/microl, while the reference range for CD8 was 145 to 884 cells/microl.

Obesity and lymphocyte subsets in virologically suppressed HIV-infected patients

The prevalence of obesity is increasing among HIVpositive patients, with current estimates—20% to 30%— rivaling estimates for the general US population [1-3]. Obesity is associated with a chronic, systemic state of inflammation that may contribute to the development of many obesity-related comorbidities. Obesity increases the risk of hypertension, diabetes, myocardial infarction, heart failure, stroke, and some malignancies [4-6]. Because adipocytes are metabolically active, obesity also increases the production of a number of substances with proinflammatory or immune-modulating functions, which might mediate the adverse health effects: adipokines, such as leptin and adiponectin; C-reactive protein; interleukin (IL)-6; tumor necrosis factor–α; and IL-1-R agonist [4,7,8] .I n a large cohort study of subjects without HIV, obesity was associated with higher CD3, CD4, and CD8 counts [9]. The treatment of morbid obesity with gastric bypass surgery in HIV-negative adults has also been reported in small studies to change cytokine levels and a number of immune markers [10,11]. In the first study, abnormal CD95 antigen expression on Tcells was reversed with surgical weight loss [10]; and in the second study, C-reactive protein levels decreased significantly postsurgery as did lymphocyte subsets CD4 and CD8 [11]. Little is known about specific effects of obesity on the immune system in HIV-positive patients. The aim of our study was to explore the effects of obesity on lymphocyte subsets in HIV-positive patients with diabetes. 2. Methods We conducted a cross-sectional study in a cohort of 216 HIV-positive patients with diabetes at the CORE Center in Chicago. The study was approved by the Cook County Bureau of Health Services institutional review board. Patients were eligible for study if they were taking highly active antiretroviral therapy (HAART) and had undetectable HIV RNA for at least 6 months. We chose to include only patients who had well-controlled HIV infection, as ongoing HIV viremia is known to cause elevations in CD8 levels. The measure of obesity was based on body mass index (BMI), calculated as weight in kilograms divided by height in meters squared: normal weight, BMI 18.5 to 24.9; overweight, BMI 25 to 29.9; obese, BMI 30 to 34.9; and morbidly obese, BMI of at least 35. Our dependent variables were the white blood cell count and the following lymphocyte subgroups, defined by surface antigens: CD3 (Tcells), CD4 (helper Tcells), CD8 (cytotoxic or suppressor T cells), CD19 (B cells), and CD56 (natural killer cells). We used linear regression models to test bivariate relationships between BMI and the concentrations of the different cell types, after visually inspecting locally weighted scatterplot smoothercurvestoensurethatlinearmodelswereappropriate. Because of consistently nonlinear relationships at BMI greater than 40, we restricted the analysis to patients with a BMI of 40 or less. We then used multivariable linear models to test the same relationships after adjusting for age, sex, ethnic group, renal function (inverse of serum creatinine), hemoglobin A1c (HbA1c) (measureofaverageglucosecontrol),andthenumber of years since HIV diagnosis. To meet model distribution assumptions,wetransformeddependent,predictor,andcontrol variables to best approximate the normal distribution (square root transformations for HIV duration and CD3, CD4, CD8, and total lymphocyte counts; log transformations for BMI and

Bronchoalveolar Lavage Fluid Eosinophils Are Correlated to Natural Killer Cells in Eosinophilic Pneumonias

Background: Eosinophilic lung diseases comprise a group of heterogeneous pulmonary disorders linked by increased eosinophils in bronchoalveolar lavage fluid (BALF). There is supporting evidence that natural killer (NK) cells participate in the regulation of eosinophilic inflammation. Objective: Our aim was to investigate the relationship between eosinophils and NK cells in BALF in patients with different interstitial lung diseases (ILDs) focusing on eosinophilic pneumonias. Methods: Of 114 patients who presented with increased BALF eosinophils (>5%), 74 patients were classified into the following groups: 27 had eosinophilic pneumonia (EP), 17 had idiopathic pulmonary fibrosis (IPF), 16 had hypersensitivity pneumonitis (HSP) and 14 had cryptogenic organizing pneumonia (COP/BOOP). Total BALF cells, cell density and cell differential counts were assessed and lymphocyte subsets CD3+, CD4+, CD8+, CD19+, CD3–CD16/56+ (NK) and CD3+CD16/56+ (NKT) were determined by flow cytometry. Results: Significant differences were observed in the percentages of lymphocytes (p < 0.001) and CD3+CD16/56+ cells (p = 0.023) among patient groups. In patients with EP, the percentage of eosinophils correlated positively with the number of CD3–CD16/56+ cells (r = 0.522, p = 0.005), the percentage of CD3–CD16/56+ cells (r = 0.690, p < 0.001), and the absolute count of CD3+CD16/56+ absolute cells (r = 0.609, p = 0.001). However, in patients with IPF, HSP or COP/BOOP, no correlation between the percentage of eosinophils and CD3–CD16/56+ or CD3+CD16/56+ cells was observed. Conclusions: Eosinophil inflammation seems to develop through a different pathway in EP compared to other ILDs.

Curative effect and histocompatibility of extracellular matrix in reconstruction of traumatic urethra defects in rabbits

Objective To study the effect and histocompatibility of urethral extracellular matrix in repair of traumatic urethra defects in rabbits. Methods Model of traumatic urethral defects was made by resecting 1.0-1.5 cm segment of the urethra in 20 rabbits. Then, the defects were repaired by a tube of extracellular matrix of the same length. The dynamic changes of CD4+, CD8+ T cell and CD4+/CD8+ in peripheral blood were detected by flow cytometry at 1,2, 3 and 4 weeks after operation. In the meantime, the immunity response of rabbits was evaluated by lymphocyte transformation test. The repaired segments stained with hematoxylin-eosin (HE) and Van Giesen were studied by histologic and pathologic observations at 10 days, 3, 6 and 24 weeks postoperatively. The urodynamics, urethroscopy and urethrography were performed at 24 weeks postoperatively. Results There was no significant difference in aspects of stimulative index of lymphocyte transformation, T lymphocyte subsets CD4+, CD8+ T cell and CD4+/CD8+ between experimental group and negative control group. Urothelium covered the whole surface of the matrix tube three weeks after operation. The smooth muscle cells increased nearly to normal urethral wall at 24 weeks. Urethrosoopy and urethrography showed glossy matrix tube. There was no statistical difference upon urodynamies between experimental group and control group. Conclusion The urethral extracellular matrix has good histocompatibility and may be a safe and effective material for repairing urethra defects. Key words: Extracellular matrix; Wounds and injuries; Urethra; Rabbits; Repair

A Phase II Study of the Purine Nucleoside Phosphorylase (PNP) Inhibitor RO5092888 (BCX-4208) in Patients with Moderate to Severe Chronic Plaque Psoriasis: Safety, Tolerability and Lymphocyte Effects

Introduction: PNP is a purine-metabolizing enzyme that catalyzes the phosphorolysis of 2′-deoxyguanosine [dGuo] to guanine and deoxyribose-1-phosphate. In T-cells, PNP inhibition leads to accumulation of deoxyguanosine triphosphate (dGTP), triggering apoptotic cell death. Chronic-plaque type psoriasis (psoriasis) is an autoimmune disorder, in which T-cells contribute, at least in part, to the manifestations and maintenance of the disease. Therefore, targeting T-cells may be a beneficial treatment strategy. Since for chronic inflammatory diseases the safety of potential immunosuppressive drugs is a major consideration, we conducted this study to investigate safety and tolerability of oral RO5092888 (BCX-4208) in patients with moderate to severe chronic plaque psoriasis.Methods: This was a randomized, double-blind, placebo-controlled, dose-ranging study. Sixty-six patients 18 to 70 years old were randomized into one of three groups: placebo, 20mg/d or 120mg/d. The study was conducted from August 2007 to June 2008. Patients received study drug over six weeks and were observed over an additional 4 weeks. Assessments for safety included tracking of adverse events (AEs) including infections; vital signs; ECGs; chemistry panel; LFTs; hematologic parameters including peripheral blood (PB) lymphocyte subsets CD3+, CD4+, CD8+, CD56+, CD20+; and urinalysis.Results: 65 of the 66 enrolled patients were analyzed. One serious AE was observed in the 20mg/d group, a deep vein thrombosis (DVT) in a patient with a history of DVTs, and was considered unrelated to study drug. The percentage of patients experiencing at least one adverse event (AE) of any grade was placebo: 33% (7/21), 20mg: 41% (9/22), and 120mg: 59% (13/22). During treatment, two infections occurred in the placebo group (influenza and sinusitis), one in the 20mg group (nasopharyngitis), and 4 in the 120mg group (2-upper respiratory tract infections, 1 bronchitis, and 1 otitis externa). Reductions in PB lymphocytes and subsets were observed (Table 1). Nine patients showed decreased levels of CD4+ lymphocytes, below 350 cells/μL (20mg/d, 3; 120mg/d, 6).Conclusion: Daily oral administration of 20 mg or 120 mg of RO5092888 for up to 6 weeks demonstrated adequate safety and tolerability. Reductions in PB T cells, T cell subsets and B cells were observed. Further investigation of RO5092888 is warranted in both T-cell and B-cell diseases.Lymphocyte subpopulationMean Nadir (% change from baseline)Placebo (n=21)20 mg (n=22)120 mg (n=22)CD3+24.030.647.5CD4+23.628.944.9CD8+26.131.853.9CD20+37.542.664.2CD56+40.147.772.6Table 1

Immunotherapy of a human papillomavirus (HPV) type 16 E7-expressing tumour by administration of fusion protein comprisingMycobacterium bovisbacille Calmette–Guérin (BCG) hsp65 and HPV16 E7

Human papillomavirus type 16 (HPV16) infection has been linked to the development of cervical and anal dysplasia and cancer. One hallmark of persistent infection is the synthesis of the viral E7 protein in cervical epithelial cells. The expression of E7 in dysplastic and transformed cells and its recognition by the immune system as a foreign antigen make it an ideal target for immunotherapy. Utilizing the E7-expressing murine tumour cell line, TC-1, as a model of cervical carcinoma, an immunotherapy based on the administration of an adjuvant-free fusion protein comprising Mycobacterium bovis BCG heat shock protein (hsp)65 linked to HPV16 E7 (hspE7) has been developed. The data show that prophylactic immunization with hspE7 protects mice against challenge with TC-1 cells and that these tumour-free animals are also protected against re-challenge with TC-1 cells. In addition, therapeutic immunization with hspE7 induces regression of palpable tumours, confers protection against tumour re-challenge and is associated with long-term survival (> 253 days). In vitro analyses indicated that immunization with hspE7 leads to the induction of a Th1-like cell-mediated immune response based on the pattern of secreted cytokines and the presence of cytolytic activity following antigenic recall. In vivo studies using mice with targeted mutations in CD8 or MHC class II or depleted of CD8 or CD4 lymphocyte subsets demonstrate that tumour regression following therapeutic hspE7 immunization is CD8-dependent and CD4-independent. These studies extend previous observations on the induction of cytotoxic T lymphocytes by hsp fusion proteins and are consistent with the clinical application of hspE7 as an immunotherapy for human cervical and anal dysplasia and cancer.

British HIV Association guidelines for HIV‐associated malignancies 2008

British HIV Association guidelines for HIV‐associated malignancies 2008

CD3-T cell receptor modulation is selectively induced in CD8 but not CD4 lymphocytes cultured in agar

The CD3-T cell receptor (TcR) complex is central to the immune response. Upon binding by specific ligands, internalized CD3-TcR molecules increase, and either T cell response or unresponsiveness may ensue depending on the triggering conditions. Using semi-solid agar culture, we have shown previously that quiescent CD4 but not CD8 lymphocytes generate clonal colonies under phytohaemagglutinin stimulation. Here we have demonstrated that the agar induces selective CD3-TcR modulation in the CD8 and not in the CD4 subset. CD8 lymphocytes preactivated in liquid culture and recultured in agar with exogenous recombinant interleukin-2 generate colonies with a modulated CD3-TcR surface expression. The peptides composing the CD3-TcR complex are synthesized in CD8 colonies as well as in CD4; however, the CD3 gamma chain is phosphorylated at a higher level in CD8 colonies. A component of the agar polymer, absent in agarose, appears to be the ligand that induces differential CD3-TcR modulation in the CD8 subset. In contrast to agar culture, CD8 colonies can be derived from quiescent CD8 lymphocytes in agarose. These CD8 colonies express unmodulated CD-TcR. CD3-TcR modulation with anti-CD3 monoclonal antibody prior to culturing in agarose inhibits the colony formation. We conclude that given triggering conditions can result in both CD3-TcR modulation and inhibition of the proliferative response selectively in the CD8 lymphocyte subset and not in the CD4.

CD4 lymphocyte subset abnormalities associated with impaired delayed cutaneous hypersensitivity reactions in patients with X-linked agammaglobulinaemia.

Circulating CD4 lymphocyte subset (CD45RA; CD45RO; CD29; Leu8) levels were determined in nine patients with X-linked agammaglobulinaemia (XLA), nine patients with common variable immunodeficiency (CVI) and in 18 age- and sex-matched controls. CD4CD45RO and CD4CD29 cells were significantly lower (P less than 0.01) in the XLA patient group (CD45RO, 15.7 +/- 10.2%; CD4CD29, 32.1 +/- 14.6%) compared with CVI patients (61.8 +/- 25.4%; 60.1 +/- 11.2%) and normal controls (43.7 +/- 22.3%, 54.5 +/- 22.0%). The levels of CD4CD45RA and CD4Leu8 cells were not abnormal in the XLA patient group. No selective reduction in CD4 subsets was observed in the CVI patient group. Delayed cutaneous hypersensitivity testing of five XLA and five CVI patients revealed a significantly reduced response to recall antigens in patients with XLA. This may relate to the deficiency of circulating memory T cells observed in these patients.

Repertoire analysis and new pathogenic epitopes of IRBP in C57BL/6 (H-2b) and B10.RIII (H-2r) mice.

Interphotoreceptor retinoid binding protein (IRBP) is the major uveitogenic retinal antigen eliciting experimental autoimmune uveoretinitis (EAU) in mice. The most frequently used mouse strains are B10.RIII and C57BL/6, but to date only one uveitogenic epitope for each has been identified. The purpose of this study was to identify and characterize additional uveitogenic epitopes in B10.RIII and C57BL/6 mice and to compare epitope recognition in wild-type versus IRBP-deficient mice on both backgrounds. Mice were immunized with IRBP. Spleen cells were stimulated in culture with overlapping peptides representing the entire IRBP molecule, and lymphocyte proliferative responses were measured. Peptides determined to be immunodominant were used to immunize mice for EAU. Cytokine profile and proliferation of the CD4 versus CD8 subsets were analyzed for the most pathogenic peptides. Two new major pathogenic epitopes were identified in WT C57BL/6 mice, residues 461-480 and 651-670. These epitopes induced EAU of severity similar to that induced by the previously known peptide, 1-20. Several other peptides elicited mild disease with lower incidence. Some peptides elicited EAU only in WT recipients of IRBP KO splenocytes. In the B10.RIII strain, two major new uveitogenic peptides were identified, 171-190 and 541-560, and several others elicited moderate disease. Unlike in C57BL/6 mice, adoptive transfer of WT B10.RIII with IRBP KO splenocytes did not reveal additional uveitogenic epitopes. Both CD4 and CD8 lymphocyte subsets proliferated to pathogenic peptides. Several new pathogenic peptides of IRBP were identified in C57BL/6 and B10.RIII mice. Differences in epitope recognition between WT and IRBP KO mice were observed in C57BL/6 mice, but not in B10.RIII mice, suggesting more extensive culling of the repertoire in C57BL/6 mice by endogenously expressed IRBP.

Peripheral Blood CD3+CD4+ and CD3−CD56+ Cell Counts and Circulating Lymphoma Cells Are Significant Predictors of Overall Survival in Newly Diagnosed Follicular Lymphoma.

BACKGROUND: Interaction between the immune system and follicular lymphoma (FL) is well recognized from immunohistochemical as well as microarray studies, but is not yet fully elucidated. A recent report suggested that a higher absolute lymphocyte count (ALC) predicted a longer time to progression and improved response to rituximab in FL (Behl et al, Br J Haem 2007). The GELA group reported a correlation between peripheral blood natural killer cell count and event free survival in diffuse large B cell lymphoma (Plonquet et al Ann Oncol 2007). The influence of lymphocyte subsets in the peripheral blood (PB) on FL outcome has not been reported.PATIENTS AND METHODS: We retrospectively analyzed the immunophenotype from peripheral blood flow cytometry (PBFC) at Fox Chase Cancer Center in FL patients (pts). We identified 127 pts (82 newly diagnosed and 45 previously treated). We recorded the presence or absence of circulating monoclonal B cells (CLC) and survival in all as well as a complete PBFC panel in > 90% of pts. FLIPI and IPI scores were calculated from data at the time of diagnosis. Overall survival and time to treatment from date of original diagnosis and from date of PBFC were analyzed in relation to ALC, CLC, the lymphocyte subsets CD3+CD4+, CD3+CD8+, CD3−CD56+/CD16+ (NK cells) and FLIPI and IPI scores. A low CD4 count was defined as <500 cells/mm3, a low NK cell count as <150 cells/mm3 and a low ALC as <1500 cells/mm3.RESULTS: There were 63 females and 64 males; median age at presentation was 56 yrs (range 31–88). Median follow up intervals from original diagnosis and from PBFC were 46 months (range 3.5 – 207) and 24 months (range 0.7–138), respectively. This abstract deals principally with the 82 newly diagnosed pts. Median follow up in the newly diagnosed subset was 27.8 months (range 0.7 to – 138); thirty-five pts were subsequently treated, and 47 pts remained under observation. FLIPI scores at diagnosis were: low - 48 pts, intermediate - 25, high - 7 and unknown - 2. IPI scores were: low - 52 pts, low-intermediate - 24, high-intermediate/high - 4 and unknown - 2. CLC were present in 21% of pts. At last follow up, 74 pts were still alive. By univariate analysis, the most significant predictors for inferior overall survival (OS) from time of PBFC were low CD4 count (P = 0.03), low NK cell count (P = 0.02), and presence of CLC (P = 0.03). There was insufficient power for significance in multivariate analysis. At last follow up, survival for the low vs. high CD4 and NK cell groups was 85% vs. 97% and 85% vs.98%, respectively. FLIPI and IPI scores, low ALC, and low CD8 did not correlate with OS from time of diagnosis or time of PBFC. With respect to time to initial treatment, only a low ALC (P<0.001) and FLIPI (P<0.001) were significant. In multivariate analysis for OS of all 127 pts, low NK cell count remained significant (P = 0.03), while low CD4 count was marginally significant (P = 0.08).CONCLUSION: Analysis of PB lymphocytes by PBFC has prognostic value in previously untreated FL pts. Low CD4 and NK cell counts as well as the presence of CLC are the most significant predictors for OS in newly diagnosed FL. Our findings suggest the importance of an intact immune system. They merit further prospective studies to ascertain the impact of treatment on subsets of pts with specific immunophenotypes and may prove useful in FL management.

Enhancement of immunological activity of CpG ODN by chitosan gene carrier

To investigate the enhancement of immunological activity of CpG ODN by chitosan gene carrier in mice, the effect of lymphocyte proliferation was detected in mice by using MTT, the levels of IgG and cytokines (IL-2 and IL-12) in serum were measured by ELISA and peripheral blood T lymphocyte subsets CD4(+), CD8(+) were analyzed by flow cytometry. Our results showed that spleen lymphocytes isolated from the CS-CpG ODN group of mice showed the strongest proliferation (SI=1.551), and the levels of IgG, IL-2 and IL-12 in serum were higher than those of other groups. Compared with the immunization with CpG ODN, the immunization with CS-CpG ODN gene carrier was more efficient in up-regulating the percentage of CD4(+)T cells and the ratio of CD4(+)/CD8(+) of mice. It was concluded that CS gene carrier of CpG ODN was much more effective in improving immunity of CpG ODN in mice.

Attenuated Disease in SIV-Infected Macaques Treated with a Monoclonal Antibody against FasL

Acute SIVmac infection in macaques is accompanied by high levels of plasma viremia that decline with the appearance of viral immunity and is a model for acute HIV disease in man. Despite specific immune responses, the virus establishes a chronic, persistent infection. The destruction of CD4+ and CD4 lymphocyte subsets in macaques contributes to viral persistence and suggests the importance of mechanisms for depleting both infected and uninfected (bystander) cells. Bystander cell killing can occur when FasL binds the Fas receptor on activated lymphocytes, which include T and B cell subpopulations that are responding to the infection. Destruction of specific immune cells could be an important mechanism for blunting viral immunity and establishing persistent infection with chronic disease. We inhibited the Fas pathway in vivo with a monoclonal antibody against FasL (RNOK203). Here we show that treatment with anti-FasL reduced cell death in circulating T and B cells, increased CTL and antibody responses to viral proteins, and lowered the setpoint viremia. By blocking FasL during only the first few weeks after infection, we attenuated SIVmac disease and increased the life span for infected and treated macaques.

Effects of Unaccustomed and Accustomed Exercise on the Immune Response in Runners

To test the hypothesis that long-term immunological response may be different after accustomed concentric and unaccustomed eccentric exercise in endurance-trained men. Fourteen highly endurance-trained male runners performed two bouts of high-intensity exercise with at least 2-wk intervals between bouts. Concentric exercise consisted of a 60-min level run with a targeted heart rate of 80% VO2 peak. Eccentric exercise was conducted lying on a specially designed exercise rack, eliciting eccentric action of the musculus quadriceps femoris. Blood samples were taken before and 1, 6, 24, 72, and 144 h after exercise to determine creatine kinase (CK), C-reactive protein (CRP), and interleukin-6 (IL-6). Lymphocyte subset distribution was assessed using flow cytometry. We found a significant (P < 0.05) increase of CD4 (eccentric: 17%; concentric: 20%), CD3+/CD4+ (16 vs 19%), CD25+ (45 vs 29%), CD25+/CD4+ (27 vs 50%), HLA-DR+ (20 vs 15%), HLA-DR+/CD4+ (16 vs 67%), and CD19+/CD45+ (52 vs 103%) positive lymphocytes 1 h after both exercise bouts. However, eccentric exercise resulted in a significantly higher and longer (6 h) increase of CD25+/CD4+ and HLA-DR+/CD8+ lymphocytes and a peak increase of CK at 72 h. IL-6 and CRP increased only after concentric exercise within the first 24 h. Both exercises resulted in a decrease of monocyte activation (LFA-1: CD18+/CD11a+) after 6 h, with an increase for the eccentric exercise part after 24 h (P < 0.05). Accustomed concentric exercise mainly induced an acute-phase response, with increased CRP, IL-6, and activation of CD4 lymphocyte subsets. Unaccustomed eccentric exercise provided a delayed increase in CK and activation of monocytes and CD4+ and CD8+ subsets. Therefore, the immunological reaction depends not only on the type of contraction but also on the adaptation to the exercise.

CD8+ natural killer cells have a potential of a sensitive and reliable biodosimetric marker in vitro

The aim of our work was to evaluate peripheral blood lymphocyte subsets as in vitro indicators of the received dose of ionizing radiation (biodosimetric markers) in the range of 3-20 Gy and to determine the appropriate time interval, during which a dose-dependent induction of apoptosis occurs upon gamma irradiation. In lymphocyte subsets characterized by double color surface immunophenotyping, four-color flow cytometry was used for visualizing cell death-associated increase in superficial phosphatidylserine exposure and cytoplasmic membrane permeability by fluorinated Annexin V and propidium iodide, respectively. No differences between sham-treated and lethal dose (7 Gy)-irradiated samples were observed upon 6 h cultivation in vitro. Ten and 18 h later, about 50 % of lymphocytes were apoptotic, but only the minority of them was in the late apoptotic phase. The only difference in radioresistance of the CD4(+)CD8(-) and CD4(-)CD8(+) lymphocyte subsets was seen upon 2-day cultivation when huge depletion of intact cells and prevalence of the late apoptotic population became obvious. A dose-dependence study in 16 and 48 h cultures confirmed the effectiveness of major T cell subsets as biodosimetric indicators. On the other hand, the minor CD8(+) subset of natural killer (NK) cells has been identified as a radiosensitive lymphocyte population the disappearance of which correlated with the received dose. We demonstrated that the CD3(-)CD8(+)NK subset can be used as a lethal/sublethal dose discriminator to 16 h cultivation. In addition, our data indicate that two-day cultivation followed by CD3/CD8 expression analysis in an intact lymphocyte population may provide a clue for low dosage biodosimetry.

Impact of Recovery of CD4 and CD8 Lymphocyte Subsets on Acute GVHD after Reduced Intensity Conditioning (RIC) Allogeneic Stem Cell Transplantation (allo-SCT).

RIC regimens for allo-CST are being explored with good results concerning feasibility and engraftment. However, little is known about the immune recovery pattern in these patients, especially the different CD4 and CD8 lymphoid T cell subsets. Here, we assessed at different time points after allo-SCT, the kinetic of recovery of naïve (CD45RA+/CD27+), central memory (CD45RA−/CD27+), and terminally differentiated (CD45RA+/CD27−) CD4+ and CD8+ T lymphocytes in 64 patients from a single center, receiving HLA-identical RIC allo-SCT. Patients and graft characteristics are: age 48 y (27–63), diagnoses: 22 myeloid malignancies (34%), 22 lymphoid malignancies (34%) and 20 metastatic solid tumors (31%). 51 pts (80%) were considered as high risk. 49 pts (77%) received a fludarabine, busulfan and ATG-based RIC, while 15 pts (23%) received a low dose irradiation-based RIC. 91% of patients received a PBSC graft, with 42 (66%) receiving CSA alone for GVHD prophylaxis and 22 (34%) receiving CSA and MMF. 24 pts (38%) developed grade 2–4 acute GVHD at a median of 49 d (26–85). In this series, in contrast to CD4+ T cell subsets, CD8+ T cell subsets had a progressive and sustained recovery in the first 3 months after allo-SCT, with acquisition of functional markers such as 2B4 and perforin. Among the different subsets analyzed, the recovery of naïve CD4+ T cells, and central memory CD4+ and CD8+ T cells, measured at day 28 after allo-SCT and before onset of grade 2–4 acute GVHD, showed a significant correlation with the risk of grade 2–4 acute GVHD (P=0.001; P=0.002 and P=0.05 respectively). Patients developing grade 2–4 acute GVHD recovered a median of 47 naïve CD4+ T cells/μL prior to onset of GVHD as compared to 11 cells/μL in patients with grade 0–1 acute GVHD (Figure). Naïve CD4+ T cell levels significantly decreased after appropriate acute GVHD treatment. In a Cox multivariate analysis taking into account all relevant risk factors for acute GVHD, early recovery of naïve CD4+/CD45RA+/CD27+ T cells in the first month after allo-SCT was the strongest parameter significantly predictive of grade 2–4 acute GVHD development (P=0.006; RR=4.0; 95%CI, 1.5–11.0). Interestingly, there was a significant correlation between the total number of CD4+ T cells infused with the allogeneic graft and the early recovery of naïve CD4+ T cells (P=0.001), suggesting that graft manipulation might represent an attractive tool towards harnessing alloreactivity after RIC-allo-SCT. [Display omitted]

Lymphocyte subsets and cytokines in women with gestational diabetes mellitus and their newborn

This study aimed to identify potential immunological markers for predicting type 1 diabetes in patients with gestational diabetes mellitus (GDM) and any immunological impairment in their newborn. In 62 GDM patients and 74 women with normal glucose tolerance (NGT), and their babies, we assessed total lymphocytes, T lymphocyte subsets CD3 and CD8 expressing T cell receptor (TCR) alpha/beta or gamma/delta, CD16 and CD19, pancreatic autoantibodies and cytokines (IL-5, IL-2, soluble receptor IL-2). At delivery, umbilical cord blood samples were taken for lymphocyte subpopulations and cytokine measurements. GDM mothers had higher levels of total lymphocytes, CD8 expressing TCR gamma/delta, and lower levels of CD3 expressing TCR alpha/beta than NGT controls. Insulin-treated GDM mothers had lower CD4 and CD4/CD8 ratios, and higher CD8 and IL-5 than diet-treated GDM or controls. Five women were positive for pancreatic autoantibodies, with lower CD4 ( p < 0.01) and CD4/CD8 ratios ( p < 0.05), and higher CD8 ( p < 0.03) and CD19 than GDM and control mothers negative for autoantibodies. GDM newborn had higher CD8 gamma/delta and lower CD16 than NGT babies. There were no significant differences in TNF-alpha concentrations in the cord blood obtained from the GDM and NGT newborn. In conclusion, GDM women and their newborn have lymphocyte subset impairments, which are more important in patients positive for autoantibodies and/or treated with insulin.

CD4 lymphopenia as a risk factor for febrile neutropenia and early death after cytotoxic chemotherapy in adult patients with cancer

Lymphopenia is frequently observed in patients with cancer and correlates with the risk of febrile neutropenia and early death after chemotherapy. The phenotype of the depleted lymphocyte populations was investigated in the current study. Peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD19, CD56) were quantified on Day 1 using fluorescence-activated cell sorting in a prospective study of 213 patients with cancer treated with chemotherapy in a single oncology ward during 12 months. Correlations between lymphocyte phenotype, clinical characteristics, and the risk of febrile neutropenia and early death within 31 days after chemotherapy were investigated in univariate and multivariate analyses. Total lymphocyte count and CD3, CD4, and CD8 lymphocyte subsets were significantly lower in patients who experienced febrile neutropenia. Total lymphocyte count and CD3, CD4, CD8, CD19, and CD56 lymphocyte subsets were significantly lower in patients who died within 31 days after chemotherapy. Using logistic regression, CD4 lymphopenia (< 450/muL; odds ratio [OR] = 2.9, 95% confidence interval [CI] = 1.5-5.9) and the dose of chemotherapy (OR = 3,9, 95% CI = 2.0-7.8) were both identified as independent risk factors for febrile neutropenia. Fifty-four percent of patients with both risk factors experienced febrile neutropenia. CD4 lymphocyte count < 450/muL was also an independent risk factor for early death (OR = 7.7, 95% CI = 1.7-35). Thirteen percent of patients with a CD4 lymphocyte count </= 450/muL died within 31 days after chemotherapy. Eighty-seven percent (14 of 16) of patients who died before Day 31 had a CD4 lymphocyte count < 450/muL. A low CD4 count was an independent risk factor for febrile neutropenia and early death in patients receiving cytotoxic chemotherapy.

Comparison of CD4 and CD8 lymphocyte counts in HIV-negative pulmonary TB patients with those in normal blood donors and the effect of antitubercular treatment: hospital-based flow cytometric study.

This study was designed to flow cytometrically determine baseline and sequential values of CD4 and CD8 lymphocyte subsets in patients without the human immunodeficiency virus and with pulmonary tuberculosis (TB) and to correlate these values with those obtained from normal male blood donors and with the radiologic extent of disease and response to therapy. We studied 39 male patients without the human immunodeficiency virus and with sputum positive for pulmonary TB who had been admitted to Military Hospital (Cardiothoracic Center) in Pune, India. Clinical, laboratory, and radiologic evaluations of these patients were done. Hematologic parameters were assessed by an automated hematology cell counter (AcT*Diff, Coulter), and T-cell subsets (CD4 and CD8) were determined flow cytometrically (EPICS-XL, Coulter). CD4 counts and percentages of CD4 were significantly lower, but CD8 values were normal, in patients with pulmonary TB when compared with values obtained in normal blood donors. The CD4/CD8 ratio was significantly lower in patients with TB. The CD4 counts normalized with antitubercular treatment. The radiologic extent of disease did not correlate well with the immune parameters studied. TB is a reversible cause of CD4 lymphocytopenia and is associated with normal numbers of CD8 cells. The radiologic extent of disease does not seem to determine the immune response.

Potential psychosocial mechanisms linking depression to immune function in elderly subjects

Although depression and immune changes in elderly subjects constitute a considerable health risk, mechanisms underlying the association between depression and immune function are unclear. The question of whether personality and social support can explain the variation in immune function during depression was addressed in 21 elderly depressive and 23 control subjects. The following variables were studied: neuroticism, extraversion, received social support, depression-related immune parameters [i.e. numbers of lymphocytes, lymphocyte subsets CD3+, CD8+, natural killer-like T cells (NKT), CD4/CD8 ratio, and interleukin-6 (Il-6)]. We found that neuroticism reduced the association between depression and Il-6 (from 62 to 22.4%) and between depression and CD3+ (from 27.6 to 21.6%), and was also directly related to Il-6 (i.e. adjusted for age and depression). Social support reduced the association between depression and NKT cells from 25 to 18%, while it was also directly related to NKT cells. Extraversion, adjusted for age and depression, was negatively related to CD4/CD8 ratio. Subjects with high extraversion and high social support had more NKT cells. We concluded that changes in immune function during depression can partly be explained by neuroticism and received social support, whereas immune function is also directly related to these psychosocial variables. Neuroticism may exert its contribution to the risk for depression partly via Il-6 production.