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PubMed2003

Rapid loss of both CD4+ and CD8+ T lymphocyte subsets during the acute phase of severe acute respiratory syndrome.

OBJECTIVE: To study the alteration of peripheral lymphocyte subsets in severe acute respiratory syndrome (SARS) patients and to help improve the early diagnosis of the disease. METHODS: Anti-coagulating blood samples from 98 SARS patients in the acute phase, 56 normal healthy blood donors, and from patients infected by HIV, CMV and EBV were collected. The T lymphocyte subsets were counted by flow cytometry using fluorescence-labeled specific monoclonal antibodies. RESULTS: A significant decrease was observed in all SARS patients in their peripheral CD4(+) and CD8(+) T lymphocyte absolute counts [256 (104) x 10(6)/L and 256 (117) x 10(6)/L, respectively], which were also lower than those of the patients infected with HIV, CMV and EBV. All patients infected with HIV, CMV and EBV had significantly higher CD8(+) T lymphocyte counts in comparison with normal controls. CONCLUSIONS: Decrease of both CD4(+) and CD8(+) T lymphocytes of patients is related to onset of SARS. T lymphocyte subset analysis would help improve the early diagnosis of the disease.

Adult↗
PubMed1999

An Italian national multicenter study for the definition of reference ranges for normal values of peripheral blood lymphocyte subsets in healthy adults.

BACKGROUND AND OBJECTIVE: Reference ranges are necessary in clinical chemistry and hematology to compare an observed value and to provide meaningful information. The aim of this multicenter study was the definition of reference ranges of the relative and absolute numbers of lymphocyte subsets by evaluating a large cohort of healthy adults and by using a standard protocol to reduce the variability in both sample preparation methodology and flow cytometer operation. Other aims of this study were the evaluation of the influence of sex, age, obesity, smoking, sport and some methodological variables on lymphocyte subsets and the comparison of differential white blood cell values obtained by flow cytometry and those obtained by hematology counters. DESIGN AND METHODS: Blood samples from 1311 healthy adults (blood donors and volunteers chosen according to the Italian law for donor selection) were analyzed to study, by flow cytometry, the immunophenotype of lymphocyte subsets and their distribution in terms of percentages and absolute values. Pre-analytical and analytical phases were performed according to the guidelines of the International Federation of Clinical Chemistry (IFCC) and the Italian Group of Cytometry (GIC). T cells were defined by the expression of CD3; T subpopulations by the coexpression of CD4 or CD8 or HLA-DR; B-lymphocytes were identified by the expression of CD19 while natural killer lymphocytes were identified by positivity of CD16 and/or CD56 without CD3. We calculated, for each laboratory and for all data collected, the frequency distribution percent values and absolute values of each lymphocyte subset. The influence of age, sex, smoking, obesity and sport was calculated by the t-test. The influence of some methodological variables was calculated by the t-test and multiple regression test. RESULTS: Fifty-three flow cytometry laboratories at different institutions in Italy participated in this study. Data was obtained from 1311 healthy adults aged from 18 to 70; 968 phenotype analyses (74%) were considered eligible for statistical analysis. Significant results were found as regards sex, smoking and some methodological variables (quantity of sample, washing procedures, brand of monoclonal antibodies and kind of instruments used). The comparison between hematology counters and cytometers showed no difference for any of the parameters considered. INTERPRETATION AND CONCLUSIONS: The large number of cases, the different kinds of laboratories and their distribution throughout the country make our sample representative of the Italian adult population. The standardization criteria of pre-analytical and analytical phases (the most important issues in evaluating reference values for an indicator) assured good reproducibility among laboratories so that the obtained reference ranges may be useful for interlaboratory comparison of results. Instruments and the brand of monoclonal antibodies may represent an inevitable cause of variability.

Blood Cells↗
PubMed1994

An analysis of lymphocyte subsets in the regional lymph nodes of patients with papillary thyroid carcinoma.

Lymphocyte subsets in the lymph nodes regional to papillary thyroid carcinoma were determined using flow cytometry to ascertain the differences in local immunological responses between elderly and young patients. Lymph nodes from age-matched patients with benign thyroid tumors were used as controls. No significant alterations in lymphocyte subsets were observed in the lymph nodes from the young patients regardless of whether metastasis was present, whereas those from the elderly patients showed significant decreases in pan T cell (CD2+, CD3+) and cytotoxic T cell (CD8+, CD8+CD11b-) populations, and a significant increase in B cells (CD19+) compared with those from both the young patients and the age-matched controls. These results indicate that local immunological alterations occur in elderly patients with papillary thyroid carcinoma, and we believe that immunological changes are one of the clinical characteristics of this tumor.

Adolescent↗
PubMed1994

Neonatal treatment with luteinizing hormone-releasing hormone analogs alters peripheral lymphocyte subsets and cellular and humorally mediated immune responses in juvenile and adult male monkeys.

We examined the effect of treatment with a LH-releasing hormone (LHRH) agonist (Ag), antagonist (Ant), or Ant and androgen (Ant/And) for the first 4 months of postnatal life on lymphocyte subsets and cellular and humorally mediated immune responses in juvenile and adult male monkeys. We also determined the effect of 9 weeks of Ant treatment on lymphocyte subsets in adult male monkeys. Adult male monkeys that had been treated neonatally with an Ag had increased levels of CD8-positive (CD8+) T-cells and reduced levels of B-cells compared to vehicle-treated controls. Lymphocytes from these animals also showed an elevated proliferative response to a variety of mitogens compared to cells from control animals. Antibody production in response to tetanus toxoid was normal in treated animals. Other neonates treated with Ant/And exhibited subnormal levels of lymphocytes, CD8+ T-cells, and B-cells at 4 months of age. Similar changes, but of lesser magnitude, were observed in animals treated with Ant alone. At 6 months of age, lymphocytes from both groups of Ant-treated monkeys exhibited an above normal proliferative response to streptolysin-O, but not to other mitogens. At 18 months of age, animals treated with Ant alone produced more antitetanus antibody in response to a tetanus toxoid booster than the controls or Ant/And-treated animals. Ant treatment was without major effect on lymphocyte subsets in adult monkeys. Serum LH and testosterone levels declined, and there was a small but significant increase in B-cells, lymphocytes expressing the interleukin-2 receptor, and the CD4+/CD8+ T-cell ratio during treatment, but these parameters normalized during the posttreatment period. The data suggest that chronic neonatal treatment with an Ag or Ant alters the development of immune system responses in male primates. The significance of these changes and their impact on the ability of these animals to respond to pathogenic agents is under investigation.

Aging↗
PubMed1991

[Lymphocyte subset reconstitution following human allogeneic bone marrow transplantation].

We analysed the reconstitution of lymphocyte subset during the first 4 weeks after human allogeneic bone marrow transplantation (BMT) in relation to the recovery of hematopoiesis. Lymphocyte subset analysis was performed with flow cytometry. We performed allogeneic BMT from HLA matched sibling donor in 9 patients. We analysed the positive percentage of each surface antigen and analysed data prior to conditioning therapy and weekly during the first 4 weeks after BMT. Results were as follows: Two or 3 weeks after BMT, percentages of CD8+ (CD8+ CD11b+) lymphocyte and CD16+ (CD16+ CD57-) lymphocyte (NK cell) were increased, and those of CD3+ lymphocyte and CD4+ (CD4+ Leu8+) lymphocyte decreased. And the ratio of CD4+ lymphocytes to CD8+ lymphocytes decreased below 1.0 at 2 or 3 weeks after BMT and remained low. In relation to the recovery of hematopoiesis, CD16+ lymphocyte (especially CD16+ CD57- lymphocyte) percentages at the third weeks correlated significantly to the recovery of granulocyte, reticulocyte, and platelet. It seems that CD16+ lymphocytes may play a role in bone marrow cell engraftment and the recovery of hematopoiesis.

Adult↗
PubMed1995

Influence of inhalational, neuroleptic and local anaesthesia on lymphocyte subset distribution.

We previously demonstrated that anaesthetics affect leukocyte response by inhibiting lymphocyte capping and metabolism and the phagocytic activity of neutrophils in some experimental models. In the present study, we investigated the effects of the clinical use of three different types of anaesthesia on lymphocyte subset distribution and activity: inhalational anaesthesia (isoflurane), a neuroleptoanalgesia, or local anaesthesia was used in patients submitted to the same operation (hysterectomy). At time 0 (before administering the anaesthesia) and at 10 min and 48 h from the start of the operation, heparinized blood was obtained from each of the 30 patients studied; three lymphocyte subpopulations were isolated (CD4+, CD8+ and CD19+). Cell number was identified for each lymphocyte subset, and (as an indication of cellular function) the intracellular contents of ATP and cAMP were evaluated by luminescence methods. The relevant results for each of the 3 anaesthetic methods were as follows: i) treatment with isoflurane induced a significant reduction in the number and function of CD4+ cells at 10 min, which was reversed at 48 h; a functional but not reversible decrease of CD19+ cells was obtained, ii) treatment with neuroleptic drugs induced a significant progressive functional impairment of CD4+ and CD19+ cells, and iii) local anaesthesia caused a significant functional impairment of CD8+ cells at 48 h and a significant functional impairment of CD19+ cells at 10 min and 48 h.

Adult↗
PubMed1999

External quality control study of lymphocyte subset tests in Japan.

We have evaluated external quality assurance of lymphocytes subset tests in Japan. The data suggest: 1) Values of lymphocyte subsets were variable at each medical laboratory in Japan. 2) Value of CD3 did not varied so much, but that of CD 19/20 or NK marker was widely varied. 3) Values of CD4 and CD8 varied in small extension. 4) CV of values was relatively low when analysis was carried out in commercial laboratories or when antibodies and instrument of Ortho-Clinical Diagnostics were used. 5) In some cases, CV was also relatively low if number of cell washing times was low or if self made lytic agents were used for cell lysing.

Antigens, CD↗
PubMed1991

Comparison between flowcytometry and immunoperoxidase staining for the enumeration of lymphocyte subsets.

Immunoperoxidase staining was compared with flowcytometry for the enumeration of lymphocyte subsets. The percentages obtained for peripheral blood lymphocytes using immunoperoxidase (CD3 = 76 CD4 = 27.9, B = 10.7 CD4/CD8 = 1.8) differed significantly from those obtained by flowcytometry (CD3 = 65.7 CD4 = 39.4, CD8 = 25.6, B = 16.7, HLA DR = 11.9 CD4/CD8 = 1.54) for certain subsets (CD3, CD4, B). There was no significant difference in lymphocyte subsets between children and adults using the same method. These differences are probably due to the different methods used to prepare lymphocytes for analysis. Other factors that should also be considered are the presence of CD4 antigen on monocytes and CD8 on natural killer cells.

Flow Cytometry↗
PubMed1993

[A study of T-lymphocyte subsets, IL-2 production and IL-2 receptor expression in peripheral blood lymphocytes in patients with nasopharyngeal carcinoma].

T lymphocyte subsets, IL-2 production and IL-2 receptor expression were detected in PBLs of 31 NPC patients and 28 normal subjects and of normal PBLs populations under the influence of serum of NPC patients. The results were as follows: CD4+ and IL-2 receptor expression, CD4+:CD8+ ratios and IL-2 production were significantly depressed and CD8+-significantly elevated in PBLs of NPC patients before radiation and of normal PBLs populations under the influence of the serum of NPC patients. CD8+ was significantly decreased and CD4+, IL-2 production and IL-2 receptor expression were depressed in patients after radiation. These suggest that cellular immunity and immunoregulation are impaired in NPC patients before and after radiation.

Adult↗
PubMed1997

Lymphocyte subset changes between 3 and 15 months of age in infants born to HIV-seropositive women in South Africa.

The evolution of T-lymphocyte subsets during infancy in perinatally HIV-infected African babies has not been previously described. In a hospital-based cohort study, T-lymphocyte subset changes were investigated in 72 South African black children born to HIV seropositive mothers. Sixteen (22.2%; children were classified as infected and 56 (77.8%) as uninfected by 18 months of age. Four (25%) of the infected infants died before the age of 9 months from HIV-related disease. The CD4 and CD8 T-lymphocyte subsets, expressed in absolute numbers, as percentages, percentiles or as ratios, were clear indicators of HIV infection at all ages between 3 and 15 months. The most marked changes were a decreased percentage of CD4 cells and an increase in percentage of CD8 cells in the infected group. In the 4 infected infants who died, CD8 count and CD4:CD8 ratio clearly predicted poor clinical outcome at 3 months. Taken together, both CD4:CD8 ratio and CD4 percentage are reliable markers of HIV infection in an African paediatric population; however, a raised CD8 lymphocyte count rather than a CD4 count is a more specific prognostic marker of disease progression in HIV infected children.

Age Factors↗
PubMed1981

Analysis of T lymphocyte subsets in cytomegalovirus mononucleosis.

Lymphocyte proliferative responses to mitogens and herpes virus antigens are diminished during cytomegalovirus (CMV) mononucleosis. We analyzed peripheral blood T lymphocytes from patients in the acute and convalescent phases of CMV mononucleosis using monoclonal antibodies directed against the T helper and the T cytotoxic-suppressor cell subsets. Acute CMV infection is associated with a reversal in the normal ratio of helper to suppressor T lymphocytes with relative and absolute decreases in T helper cells and corresponding increases in T suppressor cells. These alterations in T lymphocyte subsets are accompanied by diminished lymphocyte responses to the mitogen concanavalin A (Con A). During convalescence, helper T lymphocytes increase, suppressor T lymphocytes decrease, and Con A responses return to normal.

Acute Disease↗
PubMed2001

Monocyte-dependent cell death of T lymphocyte subsets in chronic hepatitis C.

A high incidence of viral persistence and progression to chronic hepatitis are characteristic features of hepatitis C virus (HCV) infection. Since T lymphocytes play a critical role in virus clearance, their abnormalities in undergoing apoptosis may cause the insufficient antiviral immune responses leading to persistent viral infection. Recent reports have suggested that monocyte-dependent cell death (MDCD) is a mechanism of T lymphocyte apoptosis in chronic viral infection. The current study was designed to monitor apoptosis of CD4+ and CD8+ T lymphocyte subsets in co-culture with CD14+ monocytes under the apoptotic stimuli in patients with chronic hepatitis C. Cell mortalities of CD4+ and CD8+ T lymphocyte subsets from patients exhibiting various degrees of chronic hepatitis C were increased to a greater extent when incubated with CD14+ monocyte subset in medium containing phorbol 12-myristate 13-acetate. The current observations demonstrate that MDCD of CD4+ and CD8+ T lymphocyte subsets may provide a cellular basis for immune dysfunction which results in insufficient viral clearance and progression of liver disease in chronic HCV infection.

Apoptosis↗
PubMedDate not supplied

T lymphocyte subsets and activation status in patients following liver transplantation.

Changes in T-lymphocyte subsets have previously been shown to relate to clinical events following liver transplantation and be of prognostic significance following renal transplantation. The aim of this study was to examine T lymphocyte subsets, their activation status and the mean fluorescence intensity of cell surface markers by flow cytometric analysis, in peripheral blood of patients following liver transplantation. Stable transplant patients (n=11) had a significantly higher level of activation (HLA-DR expression ) of all T cell subsets: CD3, CD4 and CD8 compared to healthy controls: 17.5% +/- 14.0 (mean +/- SD) vs 4.7 +/- 1.8 (p=0.04), 13.7% +/- 10.3 vs 4.3 +/- 1.7 (p=0.03) and 23.8% +/- 19.9 vs 3.6 +/- 2.4 (p=0.02) respectively. A further increase in activation status occurred in all T cell subsets in association with acute cellular rejection, reaching significance for the CD4+ population: 13.7% +/- 10.2 vs 23.3% +/- 20.6 (p=0.04). The mean fluorescence intensity of the CD3+DR- and CD3+ DR+ populations were increased to 1397 +/- 869 and 1282 +/- 810 following liver transplantation compared to values of 425 +/- 204 and 376 +/- 166 respectively for controls (p<0.05). T-lymphocytes maintain a high level of activation following liver transplantation and continue to express high levels of the surface marker CD3, which may account for the occurrence of acute cellular rejection despite immunosuppression in these patients.

Adult↗
PubMed1996

Differential expression of cytokine genes in CD27-positive and -negative CD4 lymphocyte subsets from healthy humans and rheumatoid arthritis patients.

Immunofluorescence analysis of CD27 expression by CD4 lymphocytes from the peripheral blood of healthy humans or rheumatoid arthritis (RA) patients and from the synovial fluid (SF) of RA patients was carried out, along with the estimation of cytokine gene [interleukin (IL) 2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-10 and interferon-gamma (IFN-gamma)] expression in these lymphocyte subsets by RT-PCR. Although no differences in CD27-positive and -negative peripheral blood CD4 cell subset distribution were revealed, marked differences in IL-3, IL-4, IL-5 and IFN-gamma mRNA expression were detected between these lymphocyte subsets and between control and disease states. These results showed that phenotyping of different cell subsets in disease cannot provide adequate information about lymphocyte functional status. To estimate differences in cytokine gene expression, CD4 lymphocytes from the peripheral blood and SF of RA patients were compared. In both cases, mRNAs for IL-2, IL-4, IL-10 and IFN-gamma were detected, but CD4 cells from SF failed to express detectable levels of IL-5 mRNA despite our findings of a CD27-cell accumulation within the synovial population of CD4 lymphocytes. These are the first data to demonstrate that expression of the IL-5 gene in RA SF CD27- lymphocytes is down-regulated and that IL-5 disregulation in RA cannot be ruled out.

Arthritis, Rheumatoid↗
PubMedDate not supplied

T lymphocyte subsets in chronic AgHBe positive hepatitis and the effects of prednisone therapy.

T lymphocyte subsets were studied in 18 patients with chronic AgHBe positive hepatitis. The study was carried out before, during and after a ten week course of prednisone therapy. There was no statistically significant difference during and after prednisone therapy in patients with hepatitis as to the total lymphocyte number and T lymphocyte subsets. The CD4/CD8 ratio was significantly decreased when compared with that of the normal controls, as the result of a reduction in CD4 T lymphocyte population before the beginning of the therapy. The patients did not respond to prednisone therapy.

CD4-CD8 Ratio↗
PubMed1999

[Development of lymphocyte subsets in tumor patients after subcutaneous administration of mistletoe extracts].

OBJECTIVE: In order to exclude the possibility that mistletoe therapy may result in immunosuppression, as indicated by a significant reduction of defined lymphocyte subsets, PATIENTS AND METHODS: peripheral blood cells of 23 tumour patients were treated subcutaneously with increasing concentrations of aqueous mistletoe extracts (Helixor(R)). RESULTS AND CONCLUSIONS: Within an observation period of 7 months, the relative amount of lymphocytes and the number of natural killer (NK) cells increased while the number of lymphocyte subsets (i. e. CD19+ B cells, CD4+ T helper cells, CD8+ CD28- suppressor cells, CD8+ CD28+ cytotoxic cells) and the proportion of CD25+ (activated) cells within T cells showed a statistically remarkable trend; due to the multiple test problem of statistical evaluation this trend is not allowed to be termed significant. The leucocytes decreased insignificantly within the observation period. However, we were unable to verify a suggested increase of defined lymphocyte subsets within 2-3 months after the onset of mistletoe treatment. Nevertheless, for the parameters CD19+ B cells, CD4+ T helper cells, CD8+ cells, CD8+ CD28+ cytotoxic cells and CD16+/CD56+ NK cells we observed statistically remarkable peaks within die 2nd and 3rd month of therapy, confirming the hypothesis. The responses to the extracts were obviously interindividually different; the immune responses especially of patients with a lower number of peripheral T cells were less significant as compared to those of patients with adequate T cell numbers. Surprisingly, even an increase of the drug concentration >3 ng mistletoe lectin (as determined within the whole plant extract) per kg body weight enhanced the number of CD4+ T helper cells. A decreased immunological reaction on mistletoe extracts was shown especially for patients with a reduced number of peripheral T cells, whereas patients with normal T-cell number were more reactive.

Adult↗
PubMed1997

Flow cytometric analysis of T-lymphocytes subsets in adult Thais.

Flow cytometer (FACScan) was used to determine the range of T lymphocyte subpopulations in normal Thai blood donors at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai. Reference population consisted of 150 healthy HIV seronegative blood donors. T lymphocyte subsets were analysed using two-color immunophenotyping of peripheral blood lymphocytes with a lysed whole blood technique and enumerated. The study showed that the normal values for CD3+ lymphocytes (percent), CD4+ lymphocytes (percent), CD8+ lymphocytes (percent), CD4/CD8 ratio, absolute CD3+ lymphocyte count, absolute CD4+ lymphocyte count and absolute CD8+ lymphocyte count were 64 +/- 8.8, 36.1 +/- 6.4, 25.7 +/- 7.3, 1.5 +/- 0.6, 1,630 +/- 600 cells/microl, 910 +/- 300 cells/microl and 670 +/- 350 cells/microl, respectively. We found that the values of CD3, CD4 and CD4/CD8 ratio were significantly lower than those in the Caucasians but those of CD8 was not significantly different. This observations have important clinical implication for the use of T lymphocyte subsets measurement, especially in the management of HIV infection in Thais. These normal ranges can be used as a reference for the decisions in clinical practice.

Adult↗
PubMed1997

[The changes of T lymphocyte subsets and serum interleukin-2 receptor in peripheral blood of the pregnant women complicated by systemic lupus erythematosus].

OBJECTIVE: To determine the changes of T lymphocyte subsets, natural killer (NK) cell and serum interleukin-2 receptor (SIL-2R) in peripheral blood of the pregnant women with systemic lupus erythematosus (SLE). METHODS: T lymphocyte subsets NK cell, and SIL-2R in peripheral blood were detected by flow cytometry (FCM) and enzyme-labeled immunosorbent assay (ELISA) in 14 cases of pregnant women complicated by SLE (SLE+NP), 18 cases of stationary phase SLE (SLE), 20 cases of normal non-pregnant women (NNP) and 20 cases of normal pregnant women (NP). RESULTS: The percentages of CD4+ cell were decreased significantly in SLE+NP group as compared with the other three groups (P < 0.01); and the ratio of CD4+/CD8+ was decreased significantly in SLE+NP group, as compared that in SLE group and in NNP group (P < 0.01). There were no difference in the number of NK cells among the four groups. The SIL-2R values were found to be increased significantly in the SLE+NP group, as compared with that of other three groups (P < 0.01). CONCLUSION: The observation on the changes of peripheral T lymphocyte subsets and SIL-2R may be helpful for monitoring progress of SLE during pregnancy.

Adult↗