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PubMed1996

Determination of gamma/delta and other T-lymphocyte subsets in bronchoalveolar lavage fluid and peripheral blood from patients with sarcoidosis and idiopathic fibrosis of the lung.

We measured five different lymphocyte subpopulations, including gamma/delta-T cells, in peripheral blood and bronchoalveolar lavage (BAL) fluid from 48 consecutive patients undergoing diagnostic BAL (patients with sarcoidosis (n = 19), patients with idiopathic pulmonary fibrosis (IPF; n = 11) and patients with other diseases of the lung). Quantitative analysis of CD3+, CD4+, CD8+, CD16/56+ cells and gamma/delta-T cells was done by flow cytometry. The proportion of CD3+ and CD4+ cells in the peripheral blood of patients with sarcoidosis and IPF was significantly diminished, while CD8+ lymphocytes and NK cells were significantly elevated compared to healthy controls. There was no significant difference for the gamma/delta-T cell subpopulation between patients with sarcoidosis, IPF and control group. The proportion of CD3+ and CD4+ cells in BAL fluid was significantly elevated in sarcoidosis compared to IPF, while CD8+ cells were significantly diminished. Natural killer cells, defined as CD16/56+ CD3- cells, showed comparable low numbers in sarcoidosis and IPF. For gamma/delta-T cells no significant difference was found between patients with sarcoidosis and IPF.

Adult↗
PubMed1999

Postnatal development of lymphocyte subsets in different compartments of the small intestine of piglets.

In contrast to rodents, all compartments of the porcine small intestine contain lymphoid cells at birth. During the first few days of life maternal antibodies and leukocytes are taken up by the intestinal epithelium. The number of intraepithelial lymphocytes (IEL) increases by a factor of 12 from Day 1 to Day 60, but in germfree pigs only a minor increase is observed. Immediately after birth, low numbers of T cells are present in the small intestinal lamina propria (LP), and many of these express neither CD4 nor CD8 on their surface. This type of subset composition is still present in germfree pigs at an age of 1.5 months. The appearance of IgA+ and IgM+ lymphocytes in the LP differs. Many more of these B cells are found in the LP of the crypts than of the villi. The development of the discrete jejunal Peyer's patches (PP) differs from that of the continuous PP in the terminal ileum. The lymphocyte subset composition shows the most obvious differences between conventional and germfree piglets and between the jejunal and ileal PP at an age of 1.5 months. Several markers, e.g. for antigen-presenting cells, memory T cells, M cells in the domes of PP, have recently become available for the pig. These should now be applied in experiments on pigs in the early postnatal period to study the mechanisms of the development of tolerance and protective immune reactions.

Animals↗
PubMed1990

Comparison of some lymphocyte markers in B-cell chronic lymphocytic leukaemia and systemic lupus erythematosus (the B lymphocyte subset).

Spontaneous mouse red-cell rosette formation (M-rosette), Leu 1 (CD 5) monoclonal antibody binding, ecto-nucleotidase enzyme reactions and mitogen responses of peripheral blood lymphocytes in 10 patients with B-cell chronic lymphocytic leukaemia (CLL), in 8 with systemic lupus erythematosus (SLE) and in 10 controls were studied. The numbers of Leu 1 (CD 5) positive B lymphocytes in CLL and SLE were higher than in controls. These are called "autoregulatory" B lymphocytes, which are thought to be independent of T-cell regulatory effects. A significant increase of mouse rosette forming cells (MRFCs) was found in CLL, while in SLE and in controls their number was low. Compared to the B lymphocytes of SLE patients and controls, those of CLL patients showed diminished responses to pokeweed mitogen stimulation parallel to their decreased level of 5'-nucleotidase. Correlations between these markers and the features of isolated M-rosette positive CLL B lymphocytes are discussed.

5'-Nucleotidase↗
PubMed2000

Kinetics of lymphocytic subsets in chicken tracheal lesions infected with infectious bronchitis virus.

The kinetics of T-cells (CD3 positive (+), CD4+ and CD8+ cells) and B-cells (IgG+, IgM+ and IgA+ cells) in chicken trachea were studied immunohistochemically and histopathologically following an intratracheal inoculation of infectious bronchitis virus (IBV). Viral antigen was detected in the cytoplasm of tracheal epithelium from 16 hr to 6 days post-inoculation (p.i.) with a peak on 4 days p.i. A few IgG+, IgM+ and IgA+ cells were detected in the submucosa from 8 hr p.i. Thereafter IgG+ and IgM+ cells were gradually increased in number, and dramatically increased from 3 days p.i., peaked on 4 days p.i., and gradually decreased after 5 days p.i. IgA+ cells were detected in a small number than IgG+ and IgM+ cells during the all experimental period. These B cells mainly existed in the lamina propria, and some cells were recognized in the interepithelial space. After 14 days p.i., small number of IgG+ and IgM+ cells were detected in the germinal center of lymph follicles in the lamina propria. From 24 to 60 hr p.i., a few number of CD3+, CD4+ and CD8+ cells were detected at the perivascular area in the lamina propria. After 3 or 4 days p.i., each positive T-cells increased rapidly in number, and reached on the peak at 5 days p.i. CD3+, CD4+ and CD8+ cells tend to distribute diffusely, perivascular area, and surrounding area of CD4+ cells, respectively. CD4+ cells were dramatically decreased from 7 days p.i., and CD3+ and CD8+ cells were decreased from 14 days p.i. No T-cells were detected in the lymph follicles in the lamina propria.

Animals↗
PubMed1990

Serial studies of evoked potentials and circulating lymphocyte subsets for multiple sclerosis: attempts to monitor progress.

A concurrent change in evoked potential measurements and quantitation of circulating T-suppressor (CD8) lymphocyte subpopulations might indicate increased subclinical disease activity. Eight untreated patients with clinically definite multiple sclerosis were monitored monthly for changes in the numbers of cells positive for CD8 markers, and hence in the ratio of CD4: CD8 positive cells. Such changes were found not to be associated with changes in evoked potentials or clinical status.

Adult↗
PubMed2006

A cytokine promoter/yellow fluorescent protein reporter transgene serves as an early activation marker of lymphocyte subsets.

A mouse containing an IL-4 promoter linked to the yellow fluorescent protein (YFP) reporter transgene was created to follow aspects of lymphocyte development and function. Following stimulation with phorbol 12-myristate 13-acetate and ionomycin, anti-CD3/CD28, antigen-specific peptide, or allogeneic cells, both CD4 and CD8 T cells expressed the transgene within 24h in a manner that was consistent with cellular activation markers. Transgene induction was inhibited by cyclosporine and FK506, suggesting that its activation occurs in an NFAT-dependent manner. B lymphocytes were also able to express the transgene when stimulated with LPS. This induction was inhibited in part by rapamycin. The results suggest that this transgene can function as an indicator of lymphocyte activation. Because YFP is not toxic and requires no preparation of the cells to view the reporter gene, this system provides a unique tool to follow lymphocyte activation in a number of model systems, such as those involving transplantation, allergy, and vaccine development.

Animals↗
PubMed2000

Kinetics of IL-2 receptor expression on lymphocyte subsets from goats infected with Mycobacterium avium subsp. paratuberculosis after specific in vitro stimulation.

Quantification of surface IL-2R expression on activated lymphocytes by flow cytometry have recently been reported to be useful in measuring cellular immunity against Mycobacterium avium subsp. paratuberculosis in goats (Whist et al., 2000, Vet. Immunol. Immunopathol. 73, 207-218). To characterise the phenotype of the peripheral lymphocytes expressing IL-2R after in vitro stimulation with purified protein derivative (PPD) from M. a. paratuberculosis, cells were processed for dual or triple colour analysis by flow cytometry (CD4 and IL-2R or CD8, gammadelta-TcR and IL-2R). To distinguish the response of antigen-specific T cells from non-specific stimulation, we performed a time-course study of proliferating cells in a group of M. a. paratuberculosis-infected animals and a control group. Following in vitro stimulation with PPD of whole blood for three different periods of time, IL-2R expression was detected mainly not only in gammadelta-T cells, but also in CD4+ and CD8+ T cells. We found a specific response of gammadelta-T cells from infected animals after 24h of stimulation. Following 120h of stimulation, however, gammadelta-T cells from control animals up-regulated IL-2R to the same level as those from infected animals, indicating either a non-specific stimulation or activation due to a first line of defence against Mycobacterium antigens. The CD4+ cells showed a specific response to PPD stimulation at all three time points. A minor population of antigen reactive gammadelta+ cells also expressed CD8. The proliferative responses differed between alphabeta and gammadelta-T cells; the IL-2R+ alphabeta T cell population mainly comprised proliferating cells, while the gammadelta+ population showed less expansion.

Animals↗
PubMed1996

Interactions of human alpha/beta and gamma/delta T lymphocyte subsets in shear flow with E-selectin and P-selectin.

We have compared the ability of human alpha/beta and gamma/delta T lymphocytes to adhere to selectin-bearing substrates, an interaction thought to be essential for homing and localization at sites of inflammation. Both T cell populations form rolling adhesions on E- and P-selectin substrates under physiologic flow conditions. Although equivalent to alpha/beta T cells in binding to E-selectin, gamma/delta T cells demonstrated greater ability to adhere to P-selectin that was purified or expressed on the surface of activated, adherent platelets. Under static conditions, 80% of gamma/delta T cells and 53% of alpha/beta T cells formed shear-resistant adhesions to P-selectin, whereas only 30% of gamma/delta and alpha/beta T cells adhered to E-selectin. The enhance ability of gamma/delta T cells to adhere to P-selectin cannot be attributed to differences in expression of the P-selectin glycoprotein ligand (PSGL-1), as all alpha/beta T cells versus approximately 75% of gamma/delta T cells expressed PSGL-1. Both cell populations expressed a similar percentage of the carbohydrate antigens sialyl LewisX and cutaneous lymphocyte-associated antigen. Depletion of lymphocyte populations or T cell clones bearing these oligosaccharides with the monoclonal antibody CSLEX-1 and HECA-452, respectively, resulted in a substantial reduction in adhesion to E-selectin and slight reduction in adhesion to P-selectin under flow conditions. Treatment of cells with an endopeptidase that selectively degrades O-sialomucins such as PSGL-1, abolished P-selectin but not E-selectin adhesion. Removal of terminal sialic acids with neuraminidase or protease treatment of cells abrogated cell adhesion to both selectin substrates. These results provide direct evidence for the presence of distinct E- and P-selectin ligands on T lymphocytes and suggest that gamma/delta T cells may be preferentially recruited to inflammatory sites during the early stages of an immune response when P-selectin is upregulated.

Antigens, CD↗
PubMedDate not supplied

Age-related changes of naive and memory CD4 rat lymphocyte subsets in mucosal and systemic lymphoid organs.

The purpose of this study was to investigate in rats, by double-label immunofluorescence and flow cytometric analysis, the age related changes in the CD4 subset of gut-associated lymphoid tissues and spleen. We found that the percentage of CD4+ T cells in Peyer's patches (PP) and spleen (SP) increased during the first 6 weeks after weaning. An age-related decrease of the CD4 subset was observed in SP of aged rats, but not in their PP. In all lymphoid tissues studied, an age-related decrease of the Thy-1+ subset was observed from weaning to 2 years of age. Analysis of the naive CD4 subset (CD45RC+) showed that in SP this subset increased during the first 9 weeks of age, and declined in aged rats. However, in PP this subset presented a slow decrease from weaning until 2 years of age. Together with the decrease of the naive subset, a sharp increase of the memory/activated CD4+ cells (CD45RC- Thy-1-) was observed in PP, and to a lesser extent in SP. When the maturation of the CD4 T cells in PP was followed during the first week after weaning, we found that an important proportion of this subset changes its phenotype at this time, from recent thymic emigrant (CD45RC- Thy-1+) to naive T cell (CD45RC+ Thy-1-) and then to activated/memory cell (CD45RC- Thy-1-). Therefore it appeared that CD4 T cells from PP mature faster than SP CD4 T cells, and they are not subject to the deleterious effect of aging. One surprising point was the different kinetics of the CD4 T cells observed in mesenteric lymph nodes (MLN). No age-related changes were observed in the CD4 subset at this site. Furthermore, the percentage of the CD45RC+ cells did not decrease in aged rats, and in the first 9 weeks of life an increase of this subset was observed.

Aging↗
PubMed2001

Amantadine in Parkinson's disease: lymphocyte subsets and IL-2 secreting T cell precursor frequencies.

The antiviral drug amantadine, that is effective in idiopathic Parkinson's disease (PD), may affect the composition and function of peripheral blood lymphocytes. In an explorative study, we therefore compared lymphocyte subpopulations and IL-2 secreting T cell precursors frequencies (HTLp-frequencies) in 15 PD patients without amantadine and six patients on long-term treatment. Five patients were investigated before and three months after the start of treatment. Group comparisons for long-term amantadine treatment showed no differences in subpopulations of B-, T-, and NK cells, and HTLp-frequencies. However, three months after initiation of treatment we noted in all five patients an increase of CD3+CD4+ and decrease of CD3+CD8+ cells, associated with an increase of the CD3+CD4+/CD3+CD8+ ratio. These changes had no effect on the HTLp-frequencies. Thus, at least for a short period of time, amantadine improves the T cell mediated immune system in PD patients.

Aged↗
PubMed1999

Effects of human immunodeficiency virus type 1 on CD4 lymphocyte subset activation.

The pathogenesis of the decline of CD4 lymphocyte counts accompanying the typical course of HIV-1 infection is not completely defined and might be related to a differential susceptibility of naive and memory cells to HIV-1 exposure. Here, we examined the effects induced by heat-inactivated HIV-1 virions on these lymphocyte populations. Exposure of CD45RA naive T cells to inactivated viral particles induced a marked decrease of both mitogenic responses and activation-induced apoptosis. Conversely, the growth of CD45RO cells was less severely restrained. Analysis of intracellular levels of cell cycle regulatory proteins revealed an arrest at the G1/S restriction point of the naive but not memory subset. This effect was associated with alterations in phosphotyrosine profile and with a marked decrease of ERK and NJK kinase activation. Finally, up-regulation of the cAMP-dependent protein kinase A (PKA) activity induced by mitogens was not affected by virus. Altogether, these findings show that interaction of HIV-1 with the T cell surface is sufficient to inhibit the proliferative response of the CD4CD45RA subset by disturbing proximal TCR signaling. This mechanism would affect renewal of naive lymphocytes, contributing in such a way to the impairment of T cell turnover during the course of HIV-1 infection.

Apoptosis↗
PubMed1995

Effect of splenectomy on T lymphocyte subsets in patients infected with the human immunodeficiency virus.

A case-control study was conducted at two institutions to determine whether the absolute CD4 lymphocyte count or the percentage of lymphocytes bearing the CD4 marker (i.e., the CD4 percentage) is a more accurate indicator of underlying immune status in splenectomized patients infected with human immunodeficiency virus (HIV). Each of nine splenectomized HIV-infected cases was matched with six nonsplenectomized HIV-infected controls--three matched for CD4 lymphocyte count and three for CD4 percentage. In analyses including the eight cases with an initial CD4 lymphocyte count of > 200/mm3, controlling for the CD4 count revealed differences between cases and controls in terms of CD4 percentage (range, 10%-41% and 17%-54%, respectively; P < .01) and Centers for Disease Control and Prevention (CDC) clinical stage (P = .06). Controlling for the CD4 percentage revealed a significant difference between cases and controls in terms of CD4 count (range, 396-1,040 and 55-784 cells/mm3, respectively; P < .01) but not CDC clinical stage (P > .7). These data suggest that the numerical relationship between the CD4 lymphocyte count and the CD4 percentage among splenectomized HIV-infected patients with more than 200 CD4 cells/mm3 differs from that among nonsplenectomized patients. The CD4 percentage appears to be a more accurate indicator of the underlying level of immune function in the former group of patients.

Adolescent↗
PubMed2000

Brief report: effect of antiretroviral agents on T-lymphocyte subset counts in healthy HIV-negative individuals. The Italian Registry on Antiretroviral Postexposure Prophylaxis.

To study the effect of antiretroviral agents on T-lymphocyte counts in HIV-negative individuals, total counts and CD4+ and CD8+ lymphocyte counts were measured in health care workers (HCW) who had been occupationally exposed to HIV who were untreated (164 HCW, group A), or had received antiretroviral postexposure prophylaxis (PEP). PEP included zidovudine (150 HCW, group B), zidovudine plus lamivudine (48 HCW, group C), or zidovudine, lamivudine, and indinavir (85 HCW, group D), at standard dosage for a mean of 30, 27, and 27 days of treatment, respectively. Lymphocyte values were collected after a mean of 44 days following exposure in group A, 48 days in group B, and 30 days both in groups C and D. Student's t-, nonparametric Mann-Whitney, and Kruskal-Wallis tests were used for statistical analysis. A slight increase in mean CD4 (range, 4.8%-6. 7%) and CD8 (range, 1.4%-9.3%) cells/mm3 was observed in each group. Gender, PEP duration, side effects, and follow-up time did not correlate with responses. Data did not vary using CD4 and CD8 percentages. These findings seem to reject any direct effects of antiretroviral agents, independent of retroviral inhibition, on proliferation and redistribution of T lymphocytes, as well as the hypothesized braking of lymphocyte apoptosis. The observed variations could reflect biologic variability.

Anti-HIV Agents↗