T cell malignancy in Richter's syndrome presenting as hyper IgM. Induction and characterization of a novel CD3+, CD4-, CD8+ T cell subset from phytohemagglutinin-stimulated patient's CD3+, CD4+, CD8+ leukemic T cells

A patient is described, having Richter's syndrome and immunodeficiency with hyper IgM, who developed suppressor T cell lymphoma (CD3+, CD4-, CD8+) following untreated helper-suppressor T cell chronic lymphocytic leukemia (CD3+, CD4+, CD8+). The neoplastic T cells in both malignancies expressed interleukin (IL) 2 receptors but were deficient in typical CD2+ and CD5+ pan T antigens. Additionally, a large percentage of malignant lymph node T cells expressed HLA-DR+ activation antigens. In vitro immunoglobulin-production experiments demonstrated that the patient's leukemic blood T cells had an excess helper function for IgM synthesis but a suppressor function for IgG and IgA synthesis by normal B and T cells. The leukemic blood T cells demonstrated a poor response to phytohemagglutinin (PHA). A defect in IL 2 receptor expression was evident in PHA-stimulated leukemic blood T cells. Of interest was the observation that PHA stimulated the induction of a novel CD3+, CD4-, CD8+ T cell subset from patient's CD3+, CD4+, CD8+ leukemic blood T cells. These PHA-induced CD3+, CD4-, CD8+ T cell subsets produced an elevated proliferative response to PHA and concanavalin A, had a helper cell function for IgM synthesis and produced highly elevated amounts of IL 2.     

Immunoregulatory CD4+ CD45R+ suppressor/inducer T lymphocyte subsets and impaired cell-mediated immunity in patients with Down''s syndrome.

The monoclonal antibodies 2H4 and 4B4 allow CD4+ and CD8+ T lymphocytes to be subdivided into CD45R+ and CDW29+ functional subpopulations. The CD4+ CD45R+ lymphocytes are designated as suppressor/inducer and CD4+ CDW29+ as helper/inducer subsets. Peripheral blood lymphocytes from 19 patients with Down's syndrome and 19 age- and sex-matched normal controls were analysed for the CD45R+ and CDW29+ subsets from the CD4+ and CD8+ T lymphocytes. The percentage of CD4+ CD45R+ cells (suppressor inducer) was markedly increased and of CD4+ CDW29+ cells (helper/inducer) decreased in all patients with Down's syndrome. In contract, the percentage of CD8+ CD45R+ and CD8+ CDW29+ subsets showed no major differences between patients with Down's syndrome and normal controls. Moreover, an alteration in the CD4+ and CD45R+ and CD4+ CDW29+ T cell subsets was accompanied by a markedly reduced proliferative response to phytohaemagglutinin and concanavalin A stimulation of the CD4+ T lymphocytes. Thus, a deficiency exists in patients with Down's syndrome in the CD4+ CDW29+ helper/inducer T cell subset which may contribute to their impaired cell-mediated immunity.     

Surface markers on human activated T lymphocytes. II. CD4 and CD8 differentiation antigens

In order to estimate the dependence of CD4 and CD8 antigen expression upon the cell activation, their presence on resting and activated human T cells was investigated. Peripheral blood lymphocytes of healthy donors derived from late (TEl) and early (TEe) E rosettes were used as resting and presumably in vivo activated T cell subsets, respectively. Expression of the above markers on T cells stimulated in vitro with PHA was also examined. It was found that both lymphocyte subsets contained similar percentage of CD4+ cells, nevertheless, TEe subset was partially enriched in CD8+ cells. PHA stimulation induced in TEe and TEl subsets the considerable increase of proportion of phenotypical opposing cells, CD8+ or CD4+, respectively. The above changes coincided with the time of the maximal DNA synthesis (the 72nd h of cell stimulation).     

In vivo dynamics of CD4-8- thymocytes. Proliferation, renewal and differentiation of different cell subsets studied by DNA biosynthetic labeling and surface antigen detection

The proliferative status of CD4-8- thymocyte subsets was determined by in vivo or in vitro labeling with bromodeoxyuridine (BrdUrd), a nonreutilized thymidine analogue detectable with a monoclonal antibody, simultaneously with relevant surface proteins. An actively cycling subset [J11d+, interleukin 2 receptor-positive (IL 2R+)] was defined, besides a relatively resting one (J11d-, Pgp1+, T cell receptor-positive). Continuous per os administration of BrdUrd showed that 85% only of CD4-8- thymocytes were labeled in 6 days confirming the existence of a relatively long-term resting subset. By contrast, CD4+8+ thymocytes were all labeled in 3-4 days. Observation of labeled CD4-8- cells after pulse labeling showed an immediate decrease of their absolute number per thymus, confirming their low autorenewal capacity. However, a small number of labeled cells which were hydroxyurea or colchicine resistant remained CD4-8- for several days and progressively acquired surface expression of IL 2R. IL 2R expression by cycling CD4-8- cells during thymus regeneration after antimitogenic drug treatment was rapid, but very transient. According to these results most CD4-8- thymocytes appear as largely engaged in a proliferation-dependent differentiative process, and do not behave as true stem cells. Consequently, this subset is principally renewed by thymic immigration of exogenously produced resting cells. However, a tenfold expansion of CD4-8- cells was found in the fetal and regenerating thymus, suggesting two proliferative phases during intrathymic CD4-8- cell maturation, the first one yielding to cell expansion and the second to cell differentiation. A tentative evaluation of daily cell immigration is proposed starting with the determination of the number of cells beginning DNA synthesis each day. A global model is finally discussed by confronting our kinetic results with the known reconstitution capacities of CD4-8- thymocyte subsets.     

Type-1 and type-2 CD8+ T-cell subsets isolated from chronic adult periodontitis tissue differ in surface phenotype and biological functions.

Cloning of CD8+ T cells expressing the alpha beta T-cell receptor from inflamed human gingiva revealed that at least two different subsets were found within the tissue and that these subsets were able to interact with each other. One subset produced high levels of interferon-gamma (IFN-gamma) and no interleukin-4 (IL-4) or IL-5, exhibited phytohaemagglutinin (PHA)- or anti-CD3-mediated cytolytic activity, and were CD28+. The other subset produced high levels of IL-4 in combination with IL-5, displayed no cytotoxicity and were CD28-. From the latter subset CD8+ T-cell clones were able to suppress the proliferative response of cytotoxic CD8+ T-cell clones. This suppression could be abolished by anti-IL-4 monoclonal antibodies. However, IL-4 alone was not able to induce the suppression. Our results indicate that CD8+ T cells might participate in local immune responses by the suppression of IFN-gamma-producing cells and by favouring humoral responses via the production of IL-4 and IL-5.     

Soluble and cellular markers of immune activation in patients with systemic sclerosis

The peripheral blood lymphocyte pattern, the lymphocyte responses in vitro, as well as the soluble markers of immune activation were studied in 24 patients with systemic sclerosis (SSc patients). The proportions of total T cells (CD3), their CD4 subset, as well as B lymphocytes were within the normal range. The relative proportion of CD8 lymphocytes, however, was significantly reduced. Patients with SSc had a slightly lower percentage of CD4/4B4+ cells, whereas their proportion of CD4/2H4+ cells was elevated as compared to healthy controls. The proportion of lymphocytes expressing the interleukin-2 receptor (IL-2R) was significantly higher in SSc patients. The proliferative responses of peripheral blood mononuclear cells to PHA stimulation were reduced in the patient group, while expression of IL-2R on lymphocytes after such in vitro stimulation was comparable to that of controls. Expression of IL-2R on patient but not control lymphocytes was increased after in vitro exposure to laminin. Such exposure failed to induce IL-2 production or cell proliferative responses. Soluble plasma IL-2R level (sIL-2R) and soluble CD8 (sCD8) molecule levels in SSc patients were significantly elevated. These results indicate the presence of an ongoing lymphocyte activation in this disease process.     

IL-7对PHA-抗CD3 LAK细胞抑制效应的研究

文章将IL 7与IL 2 ,PHA及αCD3联用诱生PHA αCD3LAK细胞 ,应用活细胞计数、MTT和APAAP等方法测定了细胞增殖活性、mIL 2R的表达、外源性IL 2的消耗、细胞毒活性和效应细胞表型等指标 ,并分析比较了IL 7的加入对以上指标的影响。结果表明IL 7的加入使PHA αCD3LAK细胞消耗IL 2的量、mIL 2R表达的量和细胞的增殖能力降低 ,细胞毒活性减弱 ,并伴有CD8+ 细胞百分率的下降。这一结果呈现出的IL 7的抑制作用尚未见报道 ,有进一步研究的价值。     

Human CD4+CD45R0+ and CD4+CD45RA+ T cells synergize in response to alloantigens.

Alloantigens, unlike recall antigens, activate both CD45RA+ (naive) and CD45R0+ (memory) CD4+ cells to the same extent. These T cell subsets may therefore interact with each other in response to alloantigens on transplanted grafts. We have investigated if the ability of activated CD4+CD45RA+ and CD4+CD45R0+ T cells to produce and respond to interleukin 2 (IL2) and IL4 may be involved in this interaction. After activation, both subsets up-regulate their IL2 receptor (IL2R) and IL4R expression, yet IL4 substantially enhanced the proliferation of the CD4+CD45RA+ but not of the CD4+CD45R0+ T cell subset, while IL2 increased the proliferation of CD4+CD45R0+ but not of the CD4+CD45RA+ T cells. Significantly, the CD4+CD45RA+ T cells synthesized two- to threefold more mRNA for IL2 than the CD4+CD45R0+ subset, while the CD4+CD45R0+ T cells synthesized mRNA for IL4 and interferon-gamma exclusively. The addition of IL2 to alloactivated CD4+CD45R0+ T cells further up-regulated their production of all three lymphokine mRNA; in contrast, IL4 induced an increase in mRNA for IL2 in only the alloactivated CD4+CD45RA+ subset. The reciprocity in the ability of both these CD4+ T cells to synthesize and respond to IL2 and IL4 may provide a rationale for the regulation of lymphokine interactions in vivo. Furthermore, the synergy between these subsets in response to alloantigens, which was directly quantitated by co-culturing CD4+CD45RA+ and CD4+CD45R0+ cells together prior to activation, may potentiate the alloreactivity against transplanted grafts in vivo.     

Stimulated proliferative responses in vertically HIV-infected children on HAART correlate with clinical and immunological markers

The objective of the study was to investigate the relationship between various CD4+ T cell subsets and the ability of peripheral blood mononuclear cells (PBMC) to proliferate to several stimuli in vertically human immunodeficiency virus type 1 (HIV-1)-infected children. We studied 29 HIV-1-infected children on highly active antiretroviral therapy (HAART) (median duration: 12.3 months). T cell subsets were determined by flow cytometry. Plasma viral load (VL) was quantified using a standardized molecular method. Proliferative responses were evaluated by [3H]-thymidine incorporation. Decreased proliferative responses of PBMC to pokeweed mitogen (PWM) were found for HIV-1-infected children in Centers for Disease Control (CDC) clinical categories B and C when compared to the control group (P < 0.05). Similarly, children with < or = 15% CD4+ T cells showed a decrease in proliferative responses to PWM (P < 0.01), anti-CD3 + anti-CD28 (P < 0.01) and phytohaemagglutinin (PHA) (P < 0.05) with respect to the control group and to children with CD4+ T cells > or = 25%. Proliferative responses to PWM, anti-CD3+, anti-CD28 and PHA had a statistically significant positive correlation with CD3+/mm3, CD4+/mm3, % CD4 T cells, CD4/CD8 ratio and the percentage of naive T cell subsets (CD4+CD45RO-HLA-DR-, CD4+ CD45RA+ CD62L+, CD4+ CD45RA+), CD4+ CD62L+ and CD4+ T cells co-expressing CD38+ (CD4+ HLA-DR-CD38+, CD4+ CD38+). Moreover, we found a negative correlation between PBMC proliferative responses and % CD8 T cells, memory, memory-activated and activated CD4+ T cell subsets. Lower proliferative responses to PWM (P < 0.01) and PHA (P < 0.01) were associated with higher VL. Our data show that higher proliferative responses to PWM, anti-CD3 + anti-CD28 and PHA are associated with both non-activated and naive CD4+ T cell subsets in HIV-1-infected children on HAART.     

Co-cultivation of CD4+ and CD8+ human T-cells leads to the appearance of CD4 cells expressing CD8 through de novo synthesis of the CD8 alpha-subunit

The appearance of a population of dual-staining CD4+CD8+ cells in human T-lymphocyte cultures has been reported by various authors, including our own observation that they are always seen in simple phytohaemagglutinin-stimulated cultures from several different donors. The purpose of the present study was to investigate factors involved in the dual-staining (DS) phenotype, and to clarify some apparent inconsistencies between published observations. Our findings can be summarised as follows. 1. A population of DS CD4+CD8+ cells always appears in PHA-stimulated T-cell cultures if they contain both CD4 and CD8 subsets. The incidence of DS cells is related to PHA concentration, but other factors are involved since DS cells are not seen in PHA-stimulated cultures of purified CD4+ or CD8+ cells. Stimulation with PHA is not a prerequisite since very similar results are seen with ConA. 2. Direct physical contact between CD4+ and CD8+ cells is required for the appearance of the DS phenotype; soluble factors alone, including IL-4, appear nor to be responsible. 3. The DS phenotype in these conditions is always CD4+ cells weakly expressing CD8 and is a consequence of de novo synthesis of the CD8alpha molecule by the CD4+ cells.     

Spontaneous lymphocyte proliferation in HTLV-I/II infection reflects preferential activation of CD8 and CD16/56 cell subsets.

Previous studies have shown that lymphocytes from HTLV-infected persons spontaneously proliferate when cultured in vitro. We investigated which cell subsets become activated in this response. Mononuclear cells from 16 HTLV-seropositive former blood donors and 9 seronegative controls were cultured for 7 days; activation was then assessed by measuring DNA synthesis in cultured cells and by monitoring CD25 expression by CD3, CD4, CD8, CD19, and CD16/56 lymphocyte subsets. Of the 16 cultures of HTLV + donor cells, 10 showed spontaneous proliferation (Prol + group) and 6 did not (Prol - group). Cytofluorometric analysis revealed a significant increase in the fractions of CD8 cells and CD16/56 cells expressing CD25 for the Prol + group, compared to the Prol- and control groups. Similarly, the fractions of CD25 cells expressing CD8 or CD16/56 were significantly increased in the Prol + group. Although neither the fraction of CD4 cells expressing CD25 nor the fraction of CD25 cells expressing CD4 were increased for the Prol + group, the modal fluorescence intensity value for CD25 expression by CD4 cells was increased, suggesting some CD4 cell activation occurred as well. Blastoid cells were, on average, 79% CD25 +, whereas the sum of CD4 + CD25 + (27%), CD8 + CD25 + (30%), CD19 + CD25 + (3%), and CD16/56 + CD25 + (35%) subsets was 95%; the presence of 17% CD8 + CD16/56 + cells accounted for most of this discrepancy. These findings indicate that spontaneous lymphocyte proliferation in HTLV infection reflects preferential activation of CD8 and CD16/56 cell subsets, apparently including the minor CD8 + CD16/56 + subset.     

Impairment of CD3-dependent and CD3-independent activation pathways in CD4+ and in CD8+ T cells from old CBA/RIJ mice

Stimulation of T cells from old mice with anti-CD3 antibodies resulted in a high variability of proliferative responses, which were 2- to 8-fold lower than the responses by T cells from young mice, even in the presence of exogenous rIL-2. Moreover, the CD4+ T cells from these old mice displayed a diminished capacity to produce IL-2 in response to anti-CD3. A partial explanation was found in the observation that T cells from the majority of old mice displayed a diminished expression of CD3 of variable intensity. However, after stimulation of the T cells with the combination of phorbol-12-myristate-13-acetate (PMA) and ionomycin to bypass CD3, 3 out of 6 old mice still exhibited 2-fold lower proliferative responses than T cells from young mice; IL-2 production by the CD4+ T cells was lower in all old mice tested. Comparison of CD4+ T cells and CD8+ T cells from old mice revealed a defective PMA/ionomycin response in both subsets, although this defect seemed more pronounced in CD4+ T cells when compared with the young counterparts. The diminished response of CD8+ T cells was accompanied by a diminished expression of the IL-2R alpha-chain. In contrast, old CD4+ T cells expressed rather higher levels of IL-2R alpha-chain than young CD4+ T cells. Altogether, multiple defects which are not necessarily the same in CD4+ and CD8+ T cells are responsible for defective T cell responses in old mice.     

Proliferation of human CD4+45R+ and CD4+45R- T helper cells is promoted by both IL-2 and IL-4 while interferon-gamma production is restricted to IL-2 activated CD4+45R- T cells

Recombinant IL-2 (rIL-2) and IL-4 (rIL-4) promote proliferation of human CD4+ T cells activated in the presence of PHA, TPA or OKT-3 monoclonal antibody (MAb), whereas the production of interferon-gamma (IFN) can be induced only by rIL-2. rIL-4 induced strong proliferative responses both in accessory cell independent assays and in the presence of autologous monocytes, but has failed to induce IFN production in any of these systems. The ability of rIL-2 to induce IFN production was strongly enhanced by the addition of monocytes, although a similar proliferative response was recorded in the absence or presence of monocytes. The MAb anti-Tac inhibited the proliferative response and the production of IFN by CD4+ T cells activated in the presence of rIL-2, whereas the proliferative response to rIL-4 was unaffected. CD4+45R+ and CD4+45R- T helper cell subsets proliferated in response to both IL-2 and IL-4. A kinetic analysis demonstrated that the production of IFN throughout a five day activation period was restricted to stimulation of CD4+45R- T cells with rIL-2. This report clearly demonstrates a dissociation of IFN production and T cell proliferation in man. While proliferation can be induced by both IL-2 and IL-4 in both the helper T cell subsets studied, IFN production was induced only in the CD4+45R- subsets and only in response to IL-2.     

Subpopulations of CD4- CD8- murine thymocytes: differences in proliferation rate in vivo and proliferative responses in vitro

Cell sorting and cytotoxic depletion procedures were used to subdivide the population of CD4- CD8- ("double-negative") thymocytes from adult CBA mice on the basis of expression of Ly-1, HSA (the "heat-stable antigen" M1/69 or B2A2), Pgp-1 glycoprotein, Thy-1, MEL-14 and the PC61 antigenic determinant on the IL2 receptor (IL2R). The level of dividing cells within these subsets was assessed by brief in vivo administration of [3H]-thymidine, followed by radioautography, or by flow cytometric cell cycle analysis after DNA staining. The capacity of the subsets to proliferate in culture, in response to stimulation with concanavalin A (Con A), or with phorbol myristate acetate (PMA) and the calcium ionophore ionomycin, was assessed in high cloning efficiency single-cell culture systems. In general, the proliferative response in culture was inversely related to the rate of cell division in vivo. Response of the double-negative subsets to Con A correlated with expression of the T cell antigen receptor complex; although a high cloning efficiency was obtained from the receptor-positive fractions, very few of the clones were cytotoxic. In particular, a major Ly-1+ HSA- Pgp-1+ double-negative subset, as well as minor Ly-1- HSA- Pgp-1+ subsets, contained very few cells in cycle in vivo, but showed a high cloning efficiency in both culture systems. Conversely, the other major double-negative subset, Ly-1- HSA+ Pgp-1-, included most of the cells in cycle, but showed a reduced cloning efficiency in response to PMA and ionomycin and failed to respond to Con A. The dividing cells within the Ly-1- HSA+ Pgp-1- group were strongly enriched in the IL2R- rather than in the IL2R+ subset, suggesting IL2 was not the growth factor maintaining their proliferation in vivo.     

Qualitative and Quantitative Analysis of T Lymphocytes During Normal Human Pregnancy

PROBLEM : Human reproduction involves contact between cells which are allogeneic to one another, however the fetus not only survives but thrives. METHODS : Aspects of T-cell-mediated immunity during normal human pregnancy were studied. PBMNCs of pregnant and nonpregnant women were stimulated with PHA and cytomegalovirus antigens (CMV). The capacity of stimulated cells to proliferate, to produce IL-2 and IFN-γ, to express IL-2 receptor (IL2R1) and the effect of rIL2 on the proliferation rate of lymphocytes were examined. FACS was utilized for T-cell subset comparisons. RESULTS : The proliferation rate, IL-2, and IFN-γ synthesis were all significantly impaired at suboptimal concentration of PHA throughout pregnancy. Exogenous rIL-2 corrected this depression of cell-mediated immunity (CMI). At optimal concentration of PHA, proliferation rate and production of IFN-γ and IL-2 were all decreased. Exogenous rIL-2 corrected these deficits only in the third trimester. Third trimester pregnant women demonstrated a significant depression of proliferation as well as IL-2 and IFN-γ production after CMV stimulation, which was partially corrected by exogenous rIL-2. FACS analysis suggested that after stimulation by CMV and optimal concentration of PHA, T cells were activated and both CD4+ and CD8+ lymphoblasts expressed normal density of IL-2R1. With suboptimal PHA, the number of activated CD4+ and CD4+IL2R1+ cells were diminished and CD4+ and CD8+ T lymphoblasts expressed lower number of IL2R1. CONCLUSIONS : CD4 T helper (Thl) cell function is down regulated progressively during the three trimesters of pregnancy without changes in the quantity of T cell subsets.     

Differential activation of cytokine genes in normal CD4-bearing T cells is stimulus dependent

Studies of cloned CD4+ T cell lines have shown that they can be separated into two distinct subsets with distinctions in their functional capabilities and by the differential release of either interleukin 2 (IL 2) (TH1/inflammatory type) or IL 4 (TH2/helper type) upon activation. To establish if in vivo-derived CD4+ T cells can exhibit distinct subsets we have investigated whether normal CD4+ T cells demonstrate differential expression of IL 2 and IL 4 mRNA, and secretion of IL 2 and IL 4 after primary stimulation in vitro. Utilizing the technique of in situ hybridization IL 2 and IL 4 gene expression in individual CD4+ T cells was readily detectable after concanavalin A (Con A) phytohemagglutinin (PHA) or pokeweed mitogen (PWM)-mediated activation. The frequencies of activated T cells producing IL 2 and IL 4 mRNA after Con A or PHA activation were approximately equivalent (30-40% of cells); however, after PWM activation the number of CD4+ T cells expressing IL 4 mRNA (78%) was more than twofold greater than the number of cells producing IL 2 mRNA (30%). Maximal levels of IL 2 gene expression occurred 24 h after mitogen activation whereas the highest levels of IL 4 mRNA were not detected until 48 h after mitogen activation. Similar distinctions in the kinetics of IL 2 and IL 4 secretion after mitogen activation were also found demonstrating good concordance in the observed expression of IL 2 and IL 4 mRNA and the levels of secreted lymphokines detected by bioassay. Most importantly, we have shown by in situ hybridization analysis that the majority of individual CD4+ T cells produce only IL 2 or IL 4 mRNA, and not both, after primary activation in vitro. By contrast, most CD4+ T cells activated in the presence of PMA and ionophore express both IL 2 and IL 4 mRNA. Our studies demonstrate that in normal, non-clonal populations of CD4+ T cells, the production of IL 2 and IL 4 is independently regulated in the majority of cells and appears to be stimulus dependent.     

Persistent human immunodeficiency virus-1 antigenaemia affects the expression of interleukin-7Ralpha on central and effector memory CD4+ and CD8+ T cell subsets

Interleukin (IL)-7 and its receptor (IL-7Ralpha) play important roles in regulating lymphopoiesis. Previous studies have reported that human immunodeficiency virus-1 (HIV-1) viraemia affects the expression of IL-7Ralpha, but its effects on CD4+ and CD8+ T cell memory subsets have not been studied. Using eight-colour flow cytometry, we compared the immunophenotypic patterns of CD4+ and CD8+ T cell subsets expressing IL-7Ralpha and activation markers, as well as circulating IL-7 levels, in three well-defined groups of HIV-1-infected subjects: successfully treated, viraemic and long-term non-progressor (LTNP). Compared with successfully treated and LTNP subjects, viraemic patients had reduced expression of IL-7Ralpha on both CD4+ and CD8+ T cells, particularly on central and effector memory T cell compartments, and substantially elevated expression of activation markers on CD8+ T cell subsets. Circulating IL-7 levels were correlated negatively with the number of CD4+ and CD8+ T cell subsets expressing IL-7Ralpha; these associations were stronger with CD4+ T cell subsets and mainly with central and effector memory cells. The expression of activation markers on CD4+ and CD8+ cell T subsets was not related to circulating IL-7 levels. A strong negative correlation was observed between central memory CD4+ or CD8+ T cells expressing IL-7Ralpha and those expressing activation markers, independently of IL-7 levels. Collectively, these results provide further insight on the role of unsuppressed viral load in disrupting the IL-7/IL-7Ralpha system and contributing to HIV-1 disease progression.     

Age-related changes of the function of T cell subsets: predominant defect of the proliferative response in CD8 positive T cell subset in aged persons

The proportion of CD8 positive cells in the peripheral blood AET-rosette forming T cells from aged persons was significantly reduced than that from young persons. The difference in the proportion between aged and young groups became more significant after proliferative response to a mitogen phytohemagglutinin (PHA) or a specific antigen tuberculin active peptide (TAP). The purified macrophage-deprived T cells (Twp), CD4 (T4) positive cells or CD8 positive cells were prepared from aged or young persons. These cell preparations lost proliferative response to PHA or TAP but showed marked proliferative response to the combined stimulation to 1 microM of ionomycin and 1 nM of phorbol-12-myristate-13-acetate (PMA) at usual culture cell density (2.5 X 10(5)/ml). Proliferative responses of these cell preparations to the combined stimulation were significantly reduced in the aged than those in the young and the magnitude of the difference in the proliferative responses between aged and young groups was more pronounced in CD8 positive cell population than in CD4 positive cell population. Although the cell preparations were relatively independent of exogenous IL-2 for the proliferative response to the combined stimulation of ionomycin and PMA at usual culture cell density, they needed exogenous IL-2 for sustained proliferation at lower culture cell density (5 X 10(3)/ml). These IL-2-dependent proliferative responses to the combined stimulation in the aged were significantly lower than those in the young and again the difference in the proliferative magnitude between aged and young groups was greater in CD8 positive population. The mechanism(s) of age-related change of the proportion and proliferative ability of T subsets were discussed.     

A Functional Study of Purified CD4+ and CD8+ Cells Isolated from Synovial Fluid of Patients with Rheumatoid Arthritis and Other Arthritides

The purpose of this investigation was to study purified synovial fluid (SF) CD4⁺ and CD8⁺ cells from patients with rheumatoid arthritis (RA) and other inflammatory joint diseases (non-RA) with respect to the proliferative response to mitogens and recombinant interleukin 2 (rIL-2). Highly purified cell subsets were isolated by an immunomagnetic technique, and spontaneous proliferation as well as proliferative responses to rIL-2 and a combination of phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) (to substitute for accessory cells) were measured. Some patients had SF CD4⁺ and/or CD8⁺ cells with moderately increased spontaneous proliferation, but only the CD4⁺ cells of the two patient groups differed significantly from the peripheral blood (PB) T-cell subsets of healthy individuals who served as controls. The response to rIL-2 was variable but generally low, although about 50% of the CD4⁺ and 20% of the SF CD8⁺ cells of both patient groups expressed the Tac antigen. The response to PHA/PMA was significantly lower for RA SF CD4⁺ cells than for non-RA SF CD4⁺ cells, which again was lower than for normal PB CD4⁺ cells. SF CD8⁺ response to PMA/PHA by both groups of patients was somewhat decreased, but not significantly lower than in the controls. Thus, the CD4⁺ cells seemed functionally more deviant than the CD8⁺ cells in both patient groups, but the abnormality was most pronounced in the RA group. The results demonstrate that the previously reported diminished response to mitogens by SF mononuclear cells is present even when SF CD4⁺ cells are cultured alone. This indicates that these T cells have a reduced response, probably because of prior activation.