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PubMed1993

[Preoperative immunotherapy in patients with gastric cancer--with special reference to analysis with monoclonal antibodies of T-lymphocyte subsets and interleukin-2 oral administration of OK-432 (Picibanil)].

The effects of oral administration of OK-432 on immune reactivities of gut associated lymphoid tissue (GALT) were studied in patients with gastric cancer. The lymphocytes of peripheral blood, regional lymph nodes and spleen in patients with gastric cancer were measured by using different monoclonal antibodies and flow cytometry. OKT3, OKT4, OKT8, OKM1, OKT4+2H4+, OKT4+2H4-, OKT8+Leu15+ and OKT8+1Leu5- were used as monoclonal antibodies. The NK activity, Interleukin-2 production and Interleukin-2 receptor were also measured. After oral administration of OK432, 1) little change was found in peripheral blood and spleen, 2) the percentages of OKT3, OKT4, OKT4+2H4- and the ratio of OKT4/OKT8 were increased in proximal regional lymph nodes, and 3) the percentages of OKT8 and OKT8+Leu15- in regional lymph nodes were not changed. These results suggest that the helper T cell activity and the antitumor immunity of regional lymph nodes were elevated by oral administration of OK-432.

Administration, Oral↗
PubMed1996

Differential expression of alpha E beta 7 integrins on bronchoalveolar lavage T lymphocyte subsets: regulation by alpha 4 beta 1-integrin crosslinking and TGF-beta.

T lymphocytes expressing the alpha E beta 7 integrin are localized and selectively retained in mucosal tissues. To investigate a potential relationship between alpha E beta 7 expression and pulmonary inflammation, the distribution of alpha E beta 7-bearing CD4+ and CD8+ T cells in peripheral blood and bronchoalveolar lavage (BAL) fluids obtained from patients with allergic asthma, sarcoidosis, hypersensitivity pneumonitis, and idiopathic pulmonary fibrosis (IPF) was determined. In contrast to the distribution in peripheral blood, BAL fluid from these patients contained high number of cells expressing alpha E beta 7 with markedly different expression patterns on CD4 or CD8 cells as well as among the various diseases. Despite similar numbers of activated CD4 cells, alpha E beta 7+CD4+ T cells ranged from 15% in asthmatics to 70% in IPF. In contrast, even in normal individuals, 60% to 90% of BAL fluid CD8+ T cells express alpha E beta 7, suggesting differential induction mechanisms on CD4 and CD8 cells. In vitro experiments revealed that a substantial proportion of peripheral blood CD+ T cells express alpha E beta 7 after stimulation with anti-CD3 antibodies, and up to 80% positive cells were found after the addition of TGF-beta. In contrast, less than 10% of CD4 cells express this particular integrin after in vitro stimulation, and the presence of TGF-beta only increased the number to 30%. Supernatants from in vitro-activated BAL cells as well as concentrated BAL fluid from patients with high alpha E beta 7 expression had no further enhancing effect. However, crosslinking of alpha 4 beta 1-, but not beta 2-integrins, significantly increased the number of alpha E beta 7 expressing CD4+ and CD8+ T cells, even in the absence of TGF-beta. These data indicate that in addition to TGF-beta, the interaction of particular T-cell subsets with specific endothelial cell and extracellular matrix proteins may upregulate alpha E beta 7 integrin expression and thereby contribute to the selective accumulation of these cells in inflammatory lung diseases.

Adult↗
PubMed1993

Differential tyrosine-specific protein phosphorylation in mouse T lymphocyte subsets. Effect of age.

We have previously reported that activation of normal murine T lymphocytes with either anti-CD3 or Con A leads to increased phosphorylation of three phosphotyrosine-containing proteins with molecular masses of 120, 80, and 40 kDa, that antibody to the alpha beta TCR heterodimer induces phosphorylation of the 120- and 80-kDa species, and that aging impairs phosphorylation of all three substrates. In our study we determine if phosphorylation of these substrates differs among T cell subsets in old and young mice. In young mice, we found that the induced levels of all three phosphorylated substrates, 10 min after addition of the mitogenic stimulus, were significantly higher in CD8 than in CD4 cells for anti-CD3 and Con A stimulation; responses to anti-TCR were also higher in CD8 than in CD4 cells for the 80 kDa, although not the 120-kDa substrate. Aging led to significant declines in the ability to phosphorylate all three substrates in the CD8 population, and also of the 80-kDa substrate in CD4 cells. Comparison of unfractionated CD4 to purified CD4 memory T cells showed that the age-dependent shift from naive to memory T cells could explain the loss with age in p80 phosphorylation, in that memory cells were significantly less able to phosphorylate p80 than were the unseparated CD4 preparations. Phosphorylation of p40 also was significantly lower in CD4 memory cells, as was phosphorylation of p120 in responses to anti-CD3. We conclude that subsets of mouse splenic T cells display different patterns of tyrosine-specific protein phosphorylation in the first 10 min after activation, and that the accumulation of memory T cells with age can account for at least some of the age-dependent decline in tyrosine protein kinase pathways.

Aging↗
PubMed1989

Lymphocyte subsets alteration in patients with argentine hemorrhagic fever.

Peripheral blood lymphocyte subpopulations were studied in 15 patients with Argentine Hemorrhagic Fever (AHF), during the acute period of the disease and in early convalescence. Anti-human Ig antibodies were used to identify B cells and monoclonal antibodies to assess T4 and T8 subsets. During the acute period of the disease, significant alterations were found in B, T4, and T8 lymphocytes (P less than .001), as well as in T4/T8 ratios (P less than .001). These abnormalities disappeared in early convalescence, around 30 days after the clinical onset. Diminished numbers of T4 lymphocytes are interpreted as relevant to the immunodepression that characterizes the acute phase of AHF.

Hemorrhagic Fever, American↗
PubMed1991

Abnormalities within CD4 and CD8 T lymphocytes subsets in type 1 (insulin-dependent) diabetes.

Abnormalities in the proportions of various T lymphocyte subpopulations have been found in a number of autoimmune diseases. Monoclonal antibodies labelled with various fluorochromes were used here to define the percentages of subsets, and especially to divide CD4+ (helper/inducer) and CD8+ (suppressor/cytotoxic) cells into phenotypic subgroups. Blood samples were analysed from 25 patients (age 10.1 +/- 3.7 years) with recently diagnosed insulin-dependent diabetes mellitus (IDDM) and 25 age- and sex-matched control subjects. The percentages of CD4+ cells and CD4+CD45RA+ cells described as naive T helper cells or suppressor/inducers were increased in the IDDM patients (P less than 0.05 and P less than 0.05. Student's t-test, respectively), whereas the percentage of CD4+CD45RA- cells (memory T-helper cells, helper/inducers) was similar in the patients and controls. The percentage of CD8+CD11b+ cells containing suppressor/effector lymphocytes was decreased in the IDDM patients as compared with the controls (P less than 0.01) but no significant difference was seen in total CD8+ cells. The percentages of CD3+ cells and the proportions of these simultaneously positive for HLA-DR antigen (activated T cells) were also increased in the recent IDDM patients (P less than 0.001 and P less than 0.05, respectively), while the proportion of CD20+ B cells was decreased (P less than 0.05). The findings support the view that disturbed immune regulation occurs in IDDM and indicate that further division of T cell subpopulations may clarify our understanding of the disease process.

Adolescent↗
PubMed1995

Ascitic fluid-associated lymphocytes in patients with metastatic ovarian cancer. Analysis of lymphocyte subsets and of phenotypical changes induced by "in vitro".

Ascitic fluid and peripheral blood samples, from seven patients affected by peritoneal carcinomatosis secondary to ovarian tumors, were analyzed to examine the phenotypic properties of ascitic fluid-associated lymphocytes (AFAL) and then asses T-cell activities in the presence of different interleukins. The AFALs showed a relative enrichment of CD2+, CD3+, CD5+, CD4+ T cells compared to the peripheral blood samples, while the contrary was true for CD56+ and CD57+ cells. The same phenomenon was observed in both CD3+, CD56+ and CD2+, CD56+ cell populations. Fewer CD71+ and CD25+ cells were found in the ascitic fluid suspensions whilst the opposite was true for HLA-DR+ lymphocytes. The data obtained from the in vitro incubation of AFALs in the presence of IL-1 (or), IL-2, IL-4, IL-6 or IFN showed that IL-1, IL-2 and IFN- can induce a two-fold increase in the percentages of HLA-DR+ T lymphocytes. IL-6, IL-2 and IL-1 seemed to have a fairly modest effect on CD25. A better survival of CD56+ cells was found only in the presence of IL-4 and IL-6. Even though preliminary and based on a limited number of patients, our results would seem to invite caution in instituting intraperitoneal administration of one or more cytokines for the treatment of such lesions in humans.

Aged↗
PubMed1984

Histometric study of the localisation of lymphocyte subsets and accessory cells in human Mantoux reactions.

Intradermal injection of purified protein derivative produced typical delayed type hypersensitivity reactions in five healthy human subjects. The major subpopulations of lymphocytes and certain accessory cells were located in frozen sections of biopsies of the lesions with monoclonal antibodies and immunohistochemical staining. The densities (expressed as number/unit area for comparison) of the different types of cells were counted at various microanatomical locations in the tissue. The inflammatory cells were concentrated in narrow zones, initially (24 h) only surrounding small blood vessels but later (48-96 h) also around sweat ducts. Lymphocytes were the predominant cell type at these sites with T4 and T8 cells randomly intermixed at a ratio similar to that in the mononuclear cell fraction of the peripheral blood samples removed at the time of biopsy. There was also a scanty diffuse infiltrate in the intervening dermis, but here the T4:T8 ratio was significantly lower than that in the peripheral blood or perivascular cuffs. There was considerable intersubject variation in the relative preponderance of T8 cells in the diffuse infiltrate. The results suggest that there is no subset selection in the initial emigration of lymphocytes through vascular endothelium in the delayed hypersensitivity reaction, but that the subsets behave differently during the subsequent migration through the tissues. It remains to be determined whether the extent to which T8 cells migrate more rapidly than T4 cells through the tissues may influence the reaction at the site of entry of organisms or antigens into the body by altering the balance of the immunoregulatory lymphocyte subsets. This may underlie some of the differences in susceptibility to infection between subjects and determine the type of granuloma that develops in a particular patient.

Adult↗
PubMed2007

The positive role of lumbar puncture in the diagnosis of Vogt-Koyanagi-Harada disease: lymphocyte subsets in the aqueous humor and cerebrospinal fluid.

PURPOSE AND METHODS: Activated T cells are believed to be important at the inflammatory sites in the pathogenesis of Vogt-Koyanagi-Harada disease. Patients are often affected by meningitis and uveitis. In this work, to examine the positive role of lumbar puncture in the diagnosis of VKH disease, we analyzed and compared the surface markers of aqueous humor cells, cerebrospinal fluid cells, and peripheral blood lymphocytes by use of flow cytometry. RESULTS: Most lymphocytes were T cells. In aqueous humor and cerebrospinal fluid, numbers of CD3(+), CD4(+), and CD4(+)CD45RO(+) cells were significantly greater than in peripheral blood lymphocytes (P < 0.01). The CD4(+)/CD8(+) ratio for aqueous humor and cerebrospinal fluid was significantly higher than that for peripheral blood cells (P < 0.01). Four weeks after starting systemic corticosteroid treatment, numbers of CD4(+) and CD4(+)CD45RO(+) cells in cerebrospinal fluid were significantly lower than before treatment. CONCLUSIONS: Profiles of surface markers were similar for cerebrospinal fluid and aqueous humor, but apparently different from that for peripheral blood cells. This suggests that cerebrospinal fluid may reflect the active local immunological reaction at sites affected by the disease, i.e. the uvea and the meninges.

Acute Disease↗
PubMed1984

Normal and tumor-bearing host macrophage factor-mediated modulation of mixed-lymphocyte reaction responsiveness: separation of T-lymphocyte subset susceptibility to enhancing and inhibitory factors.

The mixed-lymphocyte reaction reactivity of normal and tumor-bearing host (TBH) T-cell subsets was examined in response to normal and TBH macrophage (M phi) supernatants. Both inhibiting and enhancing activities were identified in normal and TBH M phi supernatants. The present data suggest that TBH M phi supernatants contained more inhibitory activity than normal host M phi supernatants and that enhancing activity of M phi supernatants was restricted to the Lyt 2,3+ population of cells. TBH Lyt 2,3+ cells were more responsive to the enhancing molecule(s) than their normal counterparts. These data were consistent with studies which implicate M phi as being partially responsible for the immune dysfunction seen in TBH, and extends previous findings on the ability of M phi to regulate the immune response in an attempt to achieve homeostasis.

Animals↗
PubMed2001

Immunophenotypic analysis of HIV-infected children: alterations within the first year of life, changes with disease progression, and longitudinal analyses of lymphocyte subsets.

Perinatal infection with human immunodeficiency virus (HIV) results in tremendous activation of the pediatric immune system. An important component of understanding the pathogenesis of this disease is to characterize and quantify antigenic indicators of activation within the peripheral lymphocyte population. We measured T-lymphocyte activation and maturation antigens in a cohort of 112 HIV-infected children treated with antiretroviral therapy according to the current standard of care. Changes in expression of CD95, HLA-DR, and CD45RO were evident in 22 HIV-infected children younger than 1 year of age. A comparison of phenotypic profiles of children in mild, moderate, and severe immune categories revealed perturbations of CD28, CD38, CD45RA, CD45RO, CD95, and HLA-DR. Finally, a novel analysis of 56 HIV-infected children based on the repeated collection of data over time (median of seven observations over 33 months) demonstrated a strong negative correlation between the percentage CD4 and the percentage of CD45RO, CD95, and HLA-DR on both CD4 and CD8 cells. Our data implicate persistent immune activation, beginning within the first year of life, as a major driving force in the pathogenesis of perinatally acquired HIV disease.

ADP-ribosyl Cyclase↗
PubMed1995

Determination of CD4 and CD8 lymphocyte subsets by a new alternative fluorescence immunoassay.

The purpose of this study was to evaluate a new alternative fluorescence immunoassay method (Zymmune CD4/CD8 Cell Monitoring Kit; Zynaxis, Inc., Malvern, Pa.) for determining the absolute CD4+ and CD8+ T-lymphocyte concentrations in whole blood. The investigation was performed as a two-site comparison of the reference whole blood flow cytometric method with the Zymmune method. In this investigation, a total of 166 patient samples were evaluated of which approximately 20% were from human immunodeficiency virus-positive individuals. The mean value for samples performed by the Zymmune CD4 assay was 1,094 (range, 74 to 2,586) cells per microliters, while the reference method yielded a mean of 890 (range, 35 to 2,033) cells per microliter. The correlation coefficient for regression analysis was 0.940. The mean value for samples performed by the Zymmune CD8 assay was 700 (range, 212 to 1,813) cells per microliter, while the reference method yielded a mean of 546 (range, 82 to 2,158) cells per microliter. The correlation coefficient for regression analysis was 0.921. No site-specific differences or trends in CD4 or CD8 values were seen when the data were analyzed by site of collection. The average precision of the CD4 assay varied from 6 to 14%, corresponding to the high and low concentration ranges. For CD8, the average precision varied from 8.3 to 16% over the respective high to low concentration ranges. We conclude that the Zymmune CD4/CD8 Cell Monitoring Kit method provides absolute CD4+ and CD8+ T-lymphocyte concentrations which are equivalent to those given by the reference flow cytometric method.

CD4-CD8 Ratio↗
PubMed2004

Different procarcinogenic potentials of lymphocyte subsets in a transgenic mouse model of chronic hepatitis B.

The immune response to hepatitis viruses is believed to be involved in the development of chronic hepatitis; however, its pathogenetic potential has not been clearly defined. The current study, using a transgenic mouse model of chronic hepatitis B, was designed to determine the relative contributions of the immune cell subsets to the progression of liver disease that induces hepatocellular carcinogenesis. Hepatitis B virus transgenic mice were adoptively transferred with CD4+ and CD8+ T cell-enriched or -depleted and B cell-depleted splenocytes obtained from hepatitis B surface antigen-primed, syngeneic nontransgenic donors. The resultant liver disease, hepatocyte apoptosis, regeneration, and tumor development were assessed and compared with the manifestations in mice that had received unfractionated spleen cells. Transfer of CD8(+)-enriched splenocytes caused prolonged disease kinetics, and a marked increase in the extent of hepatocyte apoptosis and regeneration. In 12 of 14 mice the transfer resulted in multiple hepatocellular carcinomas (HCCs) comparable with the manifestations seen in the mice transferred with total splenocytes. In contrast, mice that had received CD4(+)-enriched cells demonstrated lower levels of liver disease and developed fewer incidences of HCC (4 of 17). The experiment also revealed that all of the groups of mice complicated with HCC developed comparable mean numbers and sizes of tumors. B-cell depletion had no effect on disease kinetics in this model. Taken together, these results demonstrate that the pathogenetic events induced by CD8+ T-cell subset are primarily responsible for the induction of chronic liver disease that increases tumor incidence, suggesting their potential in triggering the process of hepatocarcinogenesis.

Animals↗