In vitro effect of interleukin-12 on antigen-specific lymphocyte proliferative responses from persons infected with human immunodeficiency virus type 1

The relationship between CD4 lymphocyte count and the in vitro effect of interleukin (IL)-12 on lymphocyte proliferative responses to Candida, tetanus toxoid, and streptokinase antigens was studied in peripheral blood mononuclear cells (PBMC from 30 human immunodeficiency virus (HIV)-infected persons and 10 seronegative controls. IL-12 significantly increased proliferative responses to microbial recall antigens of PBMC from HIV-infected persons with >200 CD4 lymphocytes/mm3 but had little effect on PBMC from patients with more advanced disease. The greatest increase was seen in patients with 200-500 CD4 cells/mm3. Results of limiting dilution analysis suggested that the increase in antigen-specific lymphocyte proliferation in the presence of IL-12 was due to an increase in the number of responding cells rather than an increase in the extent of proliferation of a fixed number of responder cells.     

Lack of intrathyroidal tumor necrosis factor alpha in Graves'' disease

Human colonic intraepithelial lymphocytes from control subjects down-regulate the proliferative responses of primed allogeneic peripheral blood mononuclear cells on rechallenge with antigens or phytohaemagglutinin (PHA). In contrast, human colonic intraepithelial lymphocytes from patients with inflammatory bowel disease fail to down-regulate the proliferative responses of primed allogeneic peripheral blood mononuclear cells on rechallenge with antigens. These findings may be important in the development and maintenance of the mucosal immunological activation of inflammatory bowel disease.     

The effects of human seminal plasma and PGE2 on mitogen induced proliferation and cytokine production of human splenic lymphocytes and peripheral blood mononuclear cells

The effects of human seminal plasma (HSP) and prostaglandin E2 (PGE2) on the proliferative responses of human splenic lymphocytes and peripheral blood mononuclear cells (PBMCs) to phytohaemagglutinin (PHA), anti-CD3 and anti-CD3 plus anti-CD28 mAb have been studied. Th1 and Th2 cytokines were also measured in the supernatants of selected cultures. Both HSP and PGE2 reproducibly inhibit the proliferative response to PHA and anti-CD3 mAb in a dose dependent manner. These effects were observed with both fresh and frozen human PBMCs and splenic lymphocytes. HSP and PGE2 however were less effective in inhibiting the co-stimulatory response induced by anti-CD3 plus anti-CD28 mAb. In addition, the HSP and PGE2 treatment used inhibited the production of the Th1 cytokines IL-2 and IFNg but had a differential modulatory effect on Th2 cytokine production, namely enhancing the production of IL-6 whilst simultaneously impairing the synthesis of IL-4 and IL-10.     

The presence of cytoplasmic lipid droplets is not sufficient to account for neutral lipid signals in the 1H MR spectra of neutrophils

Stimulation of human peripheral blood neutrophils with lipopolysaccharide (LPS), arachidonic acid (AA) and oleic acid (OA) resulted in significant increases in cytoplasmic lipid droplets. This phenomenon was also observed in enucleated and degranulated cytoplasts prepared from neutrophils stimulated with LPS. In contrast, only LPS and high concentrations of OA (10 microM) produced an increase in the lipid intensities of the MR spectra of neutrophils as determined by COSY cross peak volume measurements. Lipid intensities in cells stimulated with OA (2.5 microM) and AA (2.5 microM) and phorbol myristate acetate (20 nM) were not elevated. LPS stimulation of resting cytoplasts resulted in increased lipid droplets but not MR lipid intensities. These data suggest that while cytoplasmic lipid droplets may correlate with MR lipid intensity under some circumstances, their presence is not sufficient to account for increased neutral lipid signals.     

gamma-Glutamyl transpeptidase-dependent lipid peroxidation in isolated hepatocytes and HepG2 hepatoma cells

Gamma-glutamyltranspeptidase (GGT), a plasma membrane-bound enzyme, provides the only activity capable to effect the hydrolysis of extracellular glutathione (GSH), thus favoring the cellular utilization of its constituent amino acids. Recent studies have shown however that in the presence of chelated iron prooxidant species can be originated during GGT-mediated metabolism of GSH, and that a process of lipid peroxidation can be started eventually in suitable lipid substrates. The present study was undertaken to verify if a GGT-dependent lipid peroxidation process can be induced in the lipids of biological membranes, including living cells, and if this effect can be sustained by the GGT highly expressed at the surface of HepG2 human hepatoma cells. In rat liver microsomes (chosen as model membrane lipid substrate) exposed to GSH and ADP-chelated iron, the addition of GGT caused a marked stimulation of lipid peroxidation, which was further enhanced by the addition of the GGT co-substrate glycyl-glycine. The same was observed in primary cultures of isolated rat hepatocytes, where the lipid peroxidation process did not induce acute toxic effects. GGT-stimulation of lipid peroxidation was dependent both on the concentration of GSH and of ADP-chelated iron. In GGT-rich HepG2 human hepatoma cells, the exposure to GSH, glycyl-glycine, and ADP-chelated iron resulted in a nontoxic lipid peroxidation process, which could be prevented by means of GGT inhibitors such as acivicin and the serine-boric acid complex. In addition, by co-incubation of HepG2 cells with rat liver microsomes, it was observed that the GGT owned by HepG2 cells can act extracellularly, as a stimulant on the GSH- and iron-dependent lipid peroxidation of microsomes. The data reported indicate that the lipid peroxidation of liver microsomes and of living cells can be stimulated by the GGT-mediated metabolism of GSH. Due to the well established interactions of lipid peroxidation products with cell proliferation, the phenomenon may bear particular significance in the carcinogenic process, where a relationship between the expression of GGT and tumor progression has been envisaged.     

Colony growth of human T-lymphocyte colony forming units (TL-CFU) after X-irradiation

The effect of X-irradiation on growth of T-lymphocyte colony forming units (TL-CFU) from human peripheral blood was investigated. The results indicate that not the number of activated TL-CFU but the number of cell cycles of colony forming cells was reduced by X-irradiation. Therefore we presume that TL-CFU belong to a relatively radio-resistant cell population within the PHA-responsive lymphocytes. Kinetic studies revealed that colony growth following irradiation was delayed mainly during the phase of the first cell cycle. Preculture of the cells for 24 hours after irradiation with 1,200 R in the absence of PHA caused total inhibition of colony growth in the subsequent agar culture. In the presence of PHA no inhibition was observed. This finding appears to reflect a repair mechanism from radiation damage of lymphocytes stimulated by PHA.     

Human T-cell responses to Trichophyton tonsurans: inhibition using the serum free medium Aim V

BACKGROUND: Proteins of the fungus Trichophyton tonsurans have been shown to give strong T cell proliferative responses in vitro using lymphocytes from individuals with immediate or delayed skin tests. Furthermore, Trichophyton-specific T-cell lines produce distinct patterns of cytokine production depending upon the skin-test reactivity of the host. However, skin-test negative individuals generally give limited responses. A recent study has demonstrated dust mite specific proliferation with lymphocytes from atopic and non-atopic subjects using the serum free medium Aim V. OBJECTIVE: Compare the T-cell reactivity to Trichophyton and dust mite antigens in Aim V and media containing 10% pooled AB serum. METHODS: Peripheral blood mononuclear cells (PBMC) were separated from subjects with different skin-test reactivities and cultured either in media with serum or serum free media. RESULTS: Proliferation to two Trichophyton extracts was decreased in serum free media among subjects with either immediate or delayed hypersensitivity. Trichophyton skin-test negative subjects gave poor proliferative responses in both culture conditions. A similar decrease in proliferation in serum free media was observed in cultures with PHA and tetanus toxoid. In contrast, the majority of individuals showed increased proliferation to dust mite antigens when cultured in serum free media. Cultures in serum free medium produced less IFNgamma, IL-4, or IL-5 compared with cultures in AB medium. CONCLUSIONS: In vitro T-cell responses to the dermatophyte fungus T. tonsurans are inhibited by the serum free medium Aim V. This inhibition is seen equally with cells from individuals with delayed and immediate hypersensitivity. Furthermore, the results do not support the view that AB serum is masking T-cell responses in skin-test negative individuals.     

Human interleukin-10 can directly inhibit T-cell growth

Human interleukin-10 (IL-10) inhibits T-cell proliferation and cytokine production in the presence of monocytes. In this study, we have investigated whether IL-10 can directly inhibit T cells. Highly purified peripheral blood T cells containing less than 0.1% CD14+ cells and unresponsive to phytohemagglutinin (PHA), were growth-inhibited by IL-10 when stimulated with immobilized OKT3 monoclonal antibody (MoAb; 55.4% inhibition). This effect was neutralized by the murine MoAb 19F1 directed against human IL-10. In addition, IL-10 inhibited by 52.5% the proliferation of a human tetanus toxoid-specific T-cell clone (TM11) induced by immobilized OKT3 MoAb in the absence of antigen-presenting function. T-cell growth inhibition by IL-10 did not reflect a cytokine-induced change in the kinetics of T-cell response to immobilized OKT3 MoAb, and was observed over a wide range of cell and OKT3 MoAb concentrations. Addition of 1% to 5% monocytes to highly purified peripheral blood T cells resulted in the emergence of proliferation to PHA and to soluble OKT3 MoAb, but did not significantly affect levels of growth inhibition by IL-10 in the presence of immobilized OKT3 MoAb. Similarly, addition of 10% monocytes to the TM11 T-cell clone resulted in the emergence of proliferation in response to tetanus toxoid, but did not significantly influence growth inhibition by IL-10 in the presence of immobilized OKT3 MoAb. When stimulated with immobilized OKT3 MoAb in the absence of accessory cells, T cells secreted IL-2. Secretion of IL-2 under these conditions was inhibited by IL-10 (51.5% inhibition). Thus, IL-10 can directly inhibit growth and IL-2 production in T cells triggered by immobilized OKT3 MoAb in the absence of monocytes.     

Lymphocyte subsets and mitogen stimulation of blood lymphocytes in normal pregnancy

PROBLEM: In normal pregnancy the maternal immune system should be directed towards tolerance or suppression in order not to reject the partly foreign feto-placental unit. The aim of this investigation was to find hallmarks of systemic immunosuppression during normal pregnancy. METHODS: Five healthy primigravidae were examined during pregnancy and postpartum with flow cytometric analysis to define T and B lymphocyte subsets in peripheral blood. In addition, we studied the proliferative response of lymphocytes to mitogens or interleukin-2 (IL-2) alone or in combination with immunomodulating drugs or interleukin-4 (IL-4). The results were compared to healthy, non-pregnant women. RESULTS: During pregnancy and early puerperium we noted an immune balance in favour of suppression, as measured by increased numbers of T "helper/suppressor" (CD4+CD45RA+) and "suppressor"/effector T cells (CD8+S6F1-), and decreased numbers of T "helper/inducer" (CD4+CD29+), T "helper/memory" (CD4+CD45RO+), killer/effector T cells (CD8+S6F1+), and Natural Killer cells (CD56+), as well as decreased numbers of activated lymphocytes expressing IL-2 receptor (CD25+) and T cells expressing HLA-DR (HLA-DR+CD3+). During pregnancy, lymphocyte proliferation was impaired in autologous serum with concanavalin A (ConA), phytohemagglutinin (PHA), or IL-2. A difference in proliferative response to PHA or IL-2 between cultures with AB serum and autologous serum is suggestive of an immunosuppressor factor in serum during pregnancy. Indomethacin significantly increased lymphocyte proliferation in autologous serum with ConA, indicating PGE2 mediated suppressor activity during pregnancy. Chlorambucil and cimetidine modulated the proliferative response to ConA, indicating an alkylating agent sensitive and a histamine dependent suppressor activity during pregnancy. CONCLUSIONS: During normal pregnancy, a state of systemic suppression of the maternal immune system seems to be present.     

Evidence for the presence of a kininogen-like species in a case of total deficiency of low and high molecular weight kininogens

EBI1-ligand chemokine (ELC) is a CC chemokine constitutively expressed in various lymphoid tissues and a high-affinity functional ligand for EBI1/CCR7, a seven transmembrane G-protein-coupled receptor originally identified as an Epstein-Barr virus (EBV)-inducible gene. Here we examined chemotactic activity of ELC on peripheral blood leukocytes. ELC attracted both CD4+ and CD8+ T cells, particularly efficiently after activation with IL-2 or with phytohemagglutinin (PHA) plus IL-2, as well as CD19+ B cells, but not CD16+ NK cells, CD14+ monocytes or neutrophils. Among CD3+ T cells, ELC attracted both CD45RO- naive and CD45RO+ memory subsets. ELC also induced vigorous calcium mobilization in T cells stimulated with IL-2 with an ED50 of 3 nM. ELC fused with the secreted form of alkaline phosphatase (ELC-SEAP) specifically bound to lymphocytes and this binding was blocked only by ELC among 10 CC chemokines so far tested. Notably, lymphocytes stimulated with IL-2 or T cells expanded by PHA plus IL-2 showed much higher levels of binding than fresh lymphocytes. Consistently, CCR7 mRNA was detected in CD4+ and CD8+ T cells as well as B cells, but not in NK cells, monocytes or neutrophils, and was dramatically increased in T cells upon treatment with IL-2 or with PHA plus IL-2. Like ELC mRNA, CCR7 mRNA was expressed in various lymphoid tissues. By in situ hybridization, ELC and CCR7 mRNA were detected in the parafollicular and inner cortical regions of a lymph node, and in the parafollicular regions of an appendix. Collectively, ELC and CCR7 may be involved in the trafficking of a broad spectrum of lymphocytes, especially activated T cells, into and within various lymphoid tissues.     

Role of neutrophils and lymphocytes in inhibition of a mouse mammary adenocarcinoma engineered to release IL-2, IL-4, IL-7, IL-10, IFN-alpha, IFN-gamma, and TNF-alpha

Impressive inhibition of tumor growth has been observed after transduction of cytokine genes into tumor cells. Secreted cytokines do not affect the proliferation of a tumor directly but activate a host immune reaction strong enough to overcome its oncogenic capacity. However, the reaction mechanisms activated are difficult to interpret; because these mechanisms have been derived from experiments with different tumors, comparisons are hindered. To compare the reactive mechanisms induced by each cytokine, BALB/c mice were challenged with the parental cells of the syngeneic spontaneous mammary adenocarcinoma TSA, or with TSA cells engineered to release IL2, IL4, IL7, IL10, IFN alpha, IFN gamma, and TNF alpha, and the tumor growth area was studied histologically, ultrastructurally, and immunohistochemically. These observations were integrated with data on the growth and rejection patterns of TSA cells in mice depleted of natural killer (NK) cells, granulocytes, CD4+, or CD8+ lymphocytes. The rejection of TSA-IL2 and TSA-TNF alpha cells was associated with the massive presence of neutrophils, that of TSA-IL4 and TSA-IL7 cells with neutrophils and very small areas of colliquative necrosis, and that of TSA-IFN alpha and TSA-IL10 cells with extensive areas of ischemic-coagulative necrosis and some neutrophils. TSA-IFN gamma cells displayed a delay in growth, but were not rejected. Their growth areas comprised necrotic zones of ischemic necrosis devoid of neutrophils. The selective depletion experiments demonstrated that rejection of engineered TSA cells depends on several leukocyte populations. The weight of each population varied with the secreted cytokine, although neutrophils and CD8+ lymphocytes constantly played the major role. Employment of the same tumor line engineered with the genes of different cytokines showed that each cytokine evokes a distinct reaction and that tumor inhibition results from a complex mechanism in which neutrophils and CD8+ lymphocytes and ischemic necrosis are of primary importance.     

U-101033E (2,4-diaminopyrrolopyrimidine), a potent inhibitor of membrane lipid peroxidation as assessed by the production of 4-hydroxynonenal, malondialdehyde, and 4-hydroxynonenal--protein adducts

4-Hydroxy-2-nonenal (4-HNE) and malondialdehyde (MDA) are major lipid peroxidation products generated by free radical attack on membranes and appear to contribute to the cytotoxic effects of oxidative stress by a mechanism involving adduct formation with cellular proteins. In the present studies, we investigated the relationship between lipid peroxidation and eventual inactivation of plasma membrane proteins using a model system consisting of purified red blood cell membranes and Fe2+/EDTA. Using this system, we also analyzed the ability of a novel antioxidant, U-101033E (2,4-diaminopyrrolopyrimidine), to inhibit lipid peroxidation and associated protein damage. Our results demonstrated that significant levels of MDA and 4-HNE are generated in this model system, and that both aldehydes are capable of cross-linking membrane proteins. In addition, we used a monoclonal antibody to demonstrate the presence of 4-HNE-protein adducts in this system. The generation of 4-HNE-protein adducts closely paralleled the time course of lipid peroxidation and membrane protein cross-linking, suggesting that 4-HNE may contribute to membrane protein cross-linking. Analysis of U-101033E in this system showed that this antioxidant inhibited lipid peroxidation, prevented the appearance of 4-HNE-protein adducts, and strongly reduced membrane protein cross-linking, with an EC50 of 0.5 microM. We also show that these antioxidant effects were not due to the scavenging of superoxide anion. Thus, these studies demonstrate the potential usefulness of U-101033E for treating certain disease processes where lipid peroxidation plays a role in disease pathogenesis.     

Chemotaxis of mitogen-activated human lymphocytes and the effects of membrane-active enzymes

Following incubation with polyclonal activators such as PHA or concanavalin A, human peripheral blood lymphocytes show directional migration towards chemotactic factors such as endotoxin-activated plasma, casein or denatured proteins. This migration is inhibited by treatment of the cells with phospholipase C and sphingomyelinase C, but little affected by proteases or glycosidases. In addition, these lymphocytes migrate towards PHA and other lymphocyte activators when these reagents are used as chemoattractants at concentrations well below their mitogenic doses. They also migrate towards staphylococcal protein A. Migration towards PHA and protein A is reduced by pretreating the lymphocytes with proteases but not with phospholipase C. These results suggest two independent membrane interactions which initiate lymphocyte chemotaxis, one acting directly via the lipid bilayer, the other involving the binding of ligands to membrane proteins.     

Estrogenic xenobiotics affect the intracellular activation signal in mitogen-induced human peripheral blood lymphocytes: immunotoxicological impact

The present study was an attempt to elucidate the effect of estrogenic xenobiotics on the proliferation of mitogen-stimulated human peripheral blood lymphocyte (PBL). Our findings follow: (a) the proliferation of PBL in response to phytohemagglutinin (PHA) was mediated by protein kinase C activity, but estrogenic xenobiotics had a strong inhibitory effect on protein kinase C activity of PHA-stimulated PBL; (b) cytoplasmic extracts from PHA-stimulated PBL greatly activated DNA replication, but estrogenic xenobiotics had a strong inhibitory effect on these activities. The results suggest that the cytoplasmic signal-generating system in mitogen-treated PBL is inhibited by estrogenic xenobiotics, and that the defect occurs at all stages in the sequence of events leading to DNA synthesis and cell proliferation.     

Study of T-lymphocyte subsets of healthy and Mycobacterium avium subsp. paratuberculosis-infected cattle

The relative contributions of T-lymphocyte subsets to host defense in cattle infected with Mycobacterium avium subsp. paratuberculosis is reported. The subsets were purified with appropriate monoclonal antibodies and a magnetic bead column separation system, and their purity was verified by flow cytometry. Biological activity of each subset, expressed as lymphoproliferation and gamma interferon (IFN-gamma) production, was measured in response to phytohemagglutinin (PHA) and an M. avium antigen preparation (A-PPD). IFN-gamma was measured by antibody capture enzyme-linked immunosorbent assay. The results showed a correlation between proliferation and IFN-gamma production in response to A-PPD but not to PHA. In response to PHA, CD4+ lymphocytes were the most prolific producers of IFN-gamma. CD8+ lymphocytes produced IFN-gamma to a lesser extent, whereas gammadelta+ T lymphocytes produced little or no IFN-gamma. Differences observed between the amount of IFN-gamma produced by CD4+ versus CD8+ cells and CD4+ versus gammadelta+ cells were significant (P < 0.01), but those between peripheral blood mononuclear cells (PBMC) and CD4+ T cells were not. Similar responses to A-PPD were observed except that PBMC produced higher levels of IFN-gamma than did CD4+ T cells. These data for cattle are similar to observations made for other animal species, where CD4+ cells are the major type of T lymphocytes producing IFN-gamma. They further suggest that whatever the role gammadelta+ T cells may play in paratuberculosis, it is not likely to be mediated by IFN-gamma production.     

Inhibitory effect of somatostatin on human T lymphocytes proliferation

Somatostatin (SOM) was originally described as a growth hormone release inhibiting factor, but SOM and its specific receptors (SOM-r) have been shown to be expressed on both normal and activated T and B lymphocytes and other immunocompetent cells. In the present study we have demonstrated that SOM strongly inhibits the proliferation of human T lymphocytes when stimulated by PHA, Con A or alloantigens. However, SOM was most effective when the T cells were stimulated by an alloantigen rather than a polyclonal activator such as PHA and ConA. Moreover, SOM strongly inhibited the expression of activation markers such as CD69 and CD25 that are expressed on T lymphocytes during alloantigen stimulation. SOM also inhibited both CD28 and CD2 mediated T cell proliferation. Whereas proliferation of T cells induced by the engagement of CD3 antigen using specific mAbs was only marginally affected. Our results would support the concept that in humans SOM plays a key role in the modulation of T cell activation by interfering with the antigen-independent pathways CD2 and CD28.     

Correlation of the suppressive activity of a biological response modifier on the proliferation of peripheral blood mononuclear cells and the reduction of HIV titer

Activation of CD4+ cells is a prerequisite for infection by the human immunodeficiency virus (HIV). Thus, any agent capable of suppressing CD4+ cell proliferation could create a refractory stage that would impede viral infection. We have reported, in a previous publication, that a biological response modifier (BRM), polyantigenic immunomodulator (PAI) substantially reduces HIV-1 titer (from 20 to 100%) in peripheral mononuclear cells (PBMC) cultures with high viral titer (p24 = 10(2)-10(5) pg/ml). We are presenting data suggesting that the reported reduction in virus titer seems to be associated with a suppressive activity of PAI on the proliferation of PBMC from intravenous drug users (IVDU) infected and non-infected with HIV-1. PAI, a well characterized BRM, is a mixture of inactivated bacterial and influenza virus vaccines. PBMC from healthy donors and IVDU individuals were exposed to PAI, phytohemagglutinin (PHA), interleukin-2 (IL-2) and to combinations of PAI with either PHA or IL-2. Appropriate controls were included. 3H-thymidine pulsing was used as indicator of cell proliferation. The stimulation index and the difference between mean cpm of test sample and control were used to measure proliferative activity. There was a low proliferative response in the PBMC cultures from IVDU and HIV-1 positive patients, but it was substantially lower in the later group. When PBMC cultures from the same group of individuals were exposed to PAI, PHA and IL-2, and to the combination of either PAI plus PHA or IL-2, the response observed in the PAI treated group was uniformly lower than in the other treated cultures. Moreover, when PAI was combined with PHA, it exerted a significant reduction in the measured parameters. The effect of PAI on IL-2 activity was negligible. A suppressive effect of a PAI has been detected on the proliferation of PBMC from IVDA and HIV-1 positive individuals. This activity may be associated with the capacity of PAI to reduce HIV titers in infected PBMC cultures.     

Surface expression of adenosine deaminase in mitogen-stimulated lymphocytes

Adenosine deaminase (ADA) expression on the surface of mitogen-stimulated lymphocytes was studied by flow cytometry. The gate for lymphocytes was located by cell size (forward scatter), cytoplasmic complexity (side scatter) and by expression of the markers CD2, CD4, CD8 and CD19. After mitogenic proliferation two populations appeared, one corresponding to non-stimulated cells, and the other consisting of larger cells which showed relatively high expression of adenosine deaminase on their surface. The increase was similar to that observed for CD71 expression, and paralleled the increase in 3H-thymidine incorporation. There was a correlation between ADA and CD71 expression (r = 0.92 for phytohaemagglutinin (PHA) and 0.97 for pokeweed mitogen (PWM)). These results suggest a role for ecto-adenosine deaminase in lymphocyte proliferation and/or triggering.     

Effect of dietary vitamin E and selenium deficiency on chicken splenocyte proliferation and cell surface marker expression

Beginning at hatching, chicks were fed a Basal diet, without vitamin E or selenium (Se) or the same diet supplemented with vitamin E (100 IU/kg) and Se (0.2 ppm). The effect of these treatments on the expression of cell surface markers (CT-1a, CD3, CD4, CD8, sIgs, and Ia) defining specific thymocyte and peripheral blood leukocyte (PBL) subpopulations were examined using flow cytometric analyses. In parallel studies the effect of the dietary deficiencies on splenocyte proliferative responses to ConA or PHA stimulation was examined. The mean expression of CD3 and CT-1a per cell was increased while CD8 and CD4 expression was decreased on thymocytes from chicks fed the Basal diet. The proportion of double negative (CD4-, CD8-) thymocytes and single positive CD8+ thymocytes was significantly decreased while single positive CD4+ and double positive (CD4+, CD8+) thymocytes were significantly increased by the dietary vitamin E and Se deficiencies. The dietary deficiencies resulted in a decreased proportion of peripheral T cells and specifically decreased the number of CD4+ PBL. The proliferative response to both ConA and PHA was impaired by the vitamin E and Se dietary deficiencies. The proliferative response could be fully reconstituted but only after vitamin E and Se supplementation for periods longer than 1 week. Plasma SeGSHpx and alpha-tocopherol levels paralleled the mitogen responsiveness observed. These results support the conclusion that vitamin E and Se deficiencies may affect both the maturation of specific lymphocyte subpopulations and the functional and proliferative capabilities of the peripheral lymphocytes.