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T-cell mediated cytotoxicity play an important role in the control of human immunodeficiency virus (HIV) infection. The polyclonal cytotoxic T lymphocyte (CTL) response against target cells infected with a recombinant vaccinia virus expressing Env, Gag, Nef or reverse transcriptase (RT) proteins has been studied in four groups of individuals: acquired immune deficiency syndrome (AIDS) patients, AIDS-related complex (ARC) patients, HIV-1 seropositive subjects and seronegative controls. CTL lines have been generated by non-specific stimulation with phytohemagglutinin and interleukin-2 and target cells have been prepared from autologous B lymphocytes. CTL from asymptomatic and ARC individuals recognized most of the various proteins of HIV-1 but those from AIDS patients had very low or absent responses to the majority of proteins, with the anti-Nef cytotoxic activity decreasing first. Two of 10 AIDS patients had demonstrable recognition of Gag p24, one of RT and eight patients had no recognition of any of the proteins. The effector cells were demonstrated to be predominantly of the CD8+ phenotype, using the appropriate monoclonal antibodies. When heterologous target cells were substituted for autologous cells, the cytotoxic response was abrogated in the vast majority of cases demonstrating its human leucocyte antigen (HLA) class I restriction. Among the 10 HIV-seronegative subjects, nine had no CTL activity against the various HIV-1 proteins but one subject was able to recognize Env and RT. In the evolution of HIV infection from the seropositive stage to AIDS, CTL polyclonal activities progressively decrease, with Nef responses disappearing first, then Env and Gag p55, followed by RT and Gag p24.

Natural killer (NK) cells have been shown to play a role in the phenomenon of resistance to transplantation of allogeneic stem cells. To explore and prevent such resistance, we treated severe combined immunodeficiency mice (SCID) with anti-NK antibodies and analysed the improved engraftment of stem cells induced by this treatment. Two groups of nine SCID mice (H-2d) were compared: group A received two injections of anti-asialo GM1 rabbit antibodies (anti-NK) on days 1 and 4; group B received two injections of normal rabbit serum. All mice were injected intravenously with 7 x 10(6) fetal liver cells from B6 mice (H-2b) on day 2. One month after fetal liver cell transplantation, all mice from group A demonstrated engraftment and chimerism; at this time, donor cells accounted for more than 50% of peripheral blood mononuclear cells (PBMC). In contrast, in group B, only one mouse had 26% of donor cells among PBMC and all other mice had less than 10%. At two months, results were virtually identical in group A (over 72% of donor cells among PBMC from all mice) and slightly improved in group B (0-38% of donor cells). After the third month and continuously until the 12th month, the stability of chimerism was established in group A (over 55% of donor cells in 7 of the 9 mice) but had virtually disappeared in group B (0-2% of donor cells in all mice). Tissue analysis demonstrated the improved reconstitution of the thymus and the spleen in mice from group A. The proliferative responses of spleen cells to phytomitogens were significantly higher in all mice from group A than in any mouse from group B. Skin allografts from a third party (H-2k) were rejected within 10 days by group A mice but not by group B mice, one year after fetal liver cell transplantation. On the whole, anti-NK antibodies were able to improve engraftment, chimerism and stability of allogeneic stem cell transplants.

A decrease in natural killer (NK) cell activity is a common feature of the immune dysfunction found in patients with human immunodeficiency virus (HIV)-induced acquired immune deficiency syndrome (AIDS). The present study was aimed at exploring the NK and the lymphokine-activated killer (LAK) cell activities of lymphocytes from HIV-seropositive subjects. The in vitro production of interleukins (IL-2 and IL-10) in response to mitogens was also studied. Two groups of HIV-seropositive subjects were studied: asymptomatic and AIDS patients. Controls were normal blood donors. The NK cell activity in peripheral blood mononuclear cells (PBMC) from AIDS patients was significantly lower than that in PBMC from both HIV-seronegative subjects and asymptomatic patients. There was no significant difference between asymptomatic patients and controls. Exposure of PBMC from all three groups of individuals to an optimal dose of IL-2 in vitro enhanced LAK cell activity. At all three effector: target cell ratios, the LAK activity in AIDS patients remained below that in normal subjects. However, the proportional increase of lytic activity with IL-2 was slightly higher in AIDS patients than in HIV-seronegative subjects. The mitogen-induced production of IL-2 was especially reduced in AIDS patients. In contrast, very high levels of mitogen-induced production of IL-10 were found in the AIDS group, as compared to asymptomatic subjects or to controls. We therefore conclude that the alteration of NK cell activity occurs at an advanced stage of HIV infection, that the reduction of cytotoxic activity is partially restored by exogenous IL-2, and that decreased production of IL-2 and increased production of IL-10 may account for part of this reduction in cytotoxicity.

Human fetal thymic epithelial cells (TEC) were cultured under serum-free conditions. The TEC were analyzed for the secretion of 14C-labelled peptides and of IL-3, IL-6 IL-7, IL-8, GM-CSF, LIF, fibronectin and thymosin alpha 1 by ELISA tests. IL-3 and IL-7 were not detected from these TEC. Lack of IL-7 and presence of thymosin alpha 1 and of the surface molecule TE-4 depicts these cells as subcapsular/medullary TEC. TEC secreted constitutively IL-6, IL-8, fibronectin and thymosin alpha 1 but not GM-CSF of LIF. Stimulants included recombinant IL-1 and monoclonal antibodies to the surface adhesion molecules ICAM-1 and LAF-3. In addition, hydrocortisone (2.7 x 10(-7) M) was used to dissect secretion patterns. Recombinant IL-1, anti ICAM-1, and anti LFA-3 alone and collectively induced modest but significant increases in the secretions of 14C-labelled peptides and of IL-6 and IL-8 which were not inhibited by HC. Recombinant IL-1 but not anti ICAM-1 and anti LFA-3 induced GM-CSF and LIF. HC inhibited the secretion of GM-CSF and LIF induced by IL-1. None of the stimulants augmented the constitutive secretion of fibronectin or thymosin alpha 1 and HC inhibited thymosin alpha 1 secretion. TEC secretion of cytokines but not thymosin alpha 1 and fibronectin appear to be regulated in a more complex manner than heretofore recognized.

The anti-retrovirus cell-mediated immunity was repeatedly investigated in seven monkeys (Macaca sylvana). Four of these animals were injected with cell-free supernatants containing human immunodeficiency viruses: two monkeys received HIV1 Bru (2.5 x 10(6) cpm), two received HIV2 Rod (1.5 x 10(6) cpm). Two additional animals were injected with a cell-free supernatant containing simian immunodeficiency virus SIV/mac 251 (1.5 x 10(6) cpm) and the last animal served as control. The four macaques infected with HIV2 Rod and SIV/mac 251 seroconverted. Freshly isolated and non stimulated peripheral blood mononuclear cells from these infected macaques and from the uninfected control were repeatedly assessed for cytolytic activity. Target cells consisted of heterologous human cell lines expressing HIV1 Bru, HIV2 Rod or SIV/mac proteins. A significant cytotoxic activity, non-restricted at the major histocompatibility complex class I (MHC-I), was demonstrated in one HIV2 Rod-infected animal (F8) and in one SIV/mac 251-infected animal (M1). This last animal showed progressively diminishing cytolytic activity that was correlated with a pronounced decrease in CD4+ lymphocytes. An AIDS-like disease developed in M1, with presence of lymphadenopathy, weight loss, diarrhea and opportunistic infections. Cytotoxic activity was active against SIV and HIV2-infected target cells in an MHC-unrestricted manner; it was specific to virus-infected cells and there was cross-reactivity between HIV2 and SIV. Cytotoxic effectors appeared to be mainly CD8+ cells. This model may prove to be very useful in evaluating the capacity of candidate AIDS vaccines to elicit effective cell-mediated immune responses.

The distribution and evolution of thymic dendritic cells (DC) were studied during human ontogeny. Immunochemical techniques were used to detect S100 protein-positive cells on fetal thymus sections, at different post-fecundation (PF) ages; an image analysis of these positive cells was then carried out. Variations in the percentage of these cells in the medulla were determined according to age: the higher percentages were seen at 12 and 16 weeks PF. Dendritic cells were present at an early stage in the thymic rudiment (7 weeks PF), a finding consistent with an origin of these cells in the fetal liver, and with the possible existence of local attraction and/or maturation factors. The expression of the CD54 adhesion molecule revealed the existence of particular interactions between DC and lymphocytes in the medullary area, and at the cortico-medullar junction. Diffuse CD11a (LFA-1) expression on lymphocytes was consistent throughout gestation. The monocyte/macrophage markers (CD11b, CD14) and the reaction of non-specific esterases showed that the distribution pattern of these cells differed from the DC pattern. These ontogenic data are related to the significant role played by DC throughout the different stages of thymopoiesis.

In vitro infectivity of the MT4 lymphoid cell line with human immunodeficiency virus (HIV) has been studied in correlation with the degree of expression of the CD4 molecule at the cell surface. To modulate this CD4 expression in vitro, pre-incubation with phorbol myristate acetate (PMA) was used. The lowest CD4 expression was obtained after 1 to 5 hours. Thereafter, a partial re-expression of OKT4 was observed, e.g., when the incubation time with PMA was extended to 20 hours. Reverse transcriptase (RT) activity decreased and was delayed proportionally to the length of incubation of cells with PMA. This observation was confirmed by the comparable variation of cytopathic effects and of p24 antigen release in culture supernatants. The decrease in HIV infectivity hence correlated with that of OKT4 expression when PMA treatment did not exceed a few hours. By contrast, after extended treatment, infectivity remained decreased although OKT4 expression reappeared.

We describe a mentally retarded 12-year-old girl with ataxia in whom diagnostic evaluation for short stature revealed isolated growth hormone (GH) deficiency and multiple central nervous system (CNS) lesions. Assessment of immunologic status, performed because of the persistence of recurrent respiratory tract infections, showed associated deficiencies of IgG2-IgG4 and specific antibody response; in addition, in vitro lymphocyte response to mitogens was low, in vitro production of interleukin-2 and of IgM was absent, and natural killer activity was decreased. The possibility that association of the CNS lesions, GH deficiency and immune defects could be due to alterations of the neuro-immuno-endocrine network secondary to a disturbance of neurotransmitters induced by precocious CNS damage of a viral or ischemic nature is discussed.

Mice with severe combined immunodeficiency (CB 17 scid) received isogeneic and allogeneic fetal liver cell transplantation. Immunological reconstitution was followed by immunoglobulin production, mitogen-induced proliferation, spleen and thymus lymphoid recolonization. Scid mice injected with isogeneic fetal liver cells showed normal IgM, IgG production and subnormal Con-A induced proliferation. Lymphoid organs were gradually repopulated. Mice having received allogeneic stem cells presented normal IgM and IgG secretion. They still had no mitogen-induced lymphocyte stimulation, two months after fetal cell transplantation, despite satisfactory repopulation of spleen and thymus. Reconstitution of cell-mediated immunity may be somewhat slower following allogeneic than isogeneic stem cell transplantation.

Differentiation of spleen prothymocytes into Thy-1.2+ cells under the influence of thymosin requires transcription of DNA and translation of RNA, as demonstrated by the effects of inhibitors of DNA, RNA and protein synthesis. The lifespan of the induction message for this T cell differentiation appeared to be relatively short (30 to 60 min) in the presence of a reversible inhibitor of protein synthesis. When anisomycin, a protein synthesis inhibitor, was first introduced into the cell suspension, induction was suppressed but it was restored when actinomycin D, a RNA synthesis inhibitor, was added following anisomycin. By flow cytometry, the RNA content was slightly increased in the presence of thymosin and this increase was enhanced by the simultaneous addition of RNA and protein synthesis inhibitors.

Phenotype and functions of cells of the T lymphocyte lineage from fetal liver, thymus, spleen and bone marrow were investigated at various ages. T lymphocyte differentiation was shown to be initiated in the thymus after the 7th week of gestation. In this organ, a large number of cells with a phenotype comparable to that of children thymocytes and with a high proliferative response to phytomitogens was observed at the 14th week. The fetal liver and bone marrow never contained many T-cells and the liver was shown to be virtually devoid of any of these cells before the 13th week. Fetal spleen contained appreciable amounts of T-cells after the 13th week. Helper and suppressor activities of fetal thymocytes and splenocytes were acquired between the 12th and the 16th week, but they were never as complete nor as potent as those of adult lymphocytes. HLA antigens were detected in very low amount in lymphocytes from the various organs at the beginning of the second trimester and their expression was significantly enhanced by in vitro incubation with alpha-interferon (alpha-IFN), a procedure that permits easier HLA typing of fetal cells.

Genetically hypertensive (LH), normotensive (LN), and low blood pressure (LL) rats of the Lyon strains have been simultaneously selected according to their systolic blood pressure (SBP). SBP and immunological parameters were investigated in 5 week-old rats. SBP reached 99 +/- 2 mmHg in LL, 108 +/- 2 mmHg in LN and 122 +/- 4 mmHg in LH. White blood cell and lymphocyte counts, thymus and spleen histology, percentages of T cells and subsets of T cells in these organs were not modified by genetic hypertension. The blastogenic responses to mitogens (Con A and PWM) of the LH rat thymocytes were similar to those observed in LN or LL rats. By contrast, the proliferative response of splenocytes to PWM was positively correlated (r = 0.93) with SBP level. In additional experiments we demonstrated that, besides the immune defect induced by neonatal thymectomy, the removal of the thymus at birth prevented the spontaneous increase of SBP in LH strains (102 +/- 2 mmHg instead of 122 +/- 4 mmHg), while only a slight decrease or no alteration was seen in LN and LL thymectomized rats. This antihypertensive effect tended to maintain SBP at a common level in the 3 strains. These results suggest that the thymus may be involved in the development of genetic hypertension.

Fetal liver transplantation has been shown to induce hematological and immunological reconstitution in irradiated rodents, dogs, horses, and sheep. Engraftment and reconstitution without GvHD has been readily obtained using histocompatible donors. When mismatched fetal donors were used, a comparatively larger number of donor cells was required, in addition to pre-treatment of host with higher doses of irradiation or irradiation plus chemotherapy. Stem cell suspensions devoid of any T lymphocyte can be transplanted across major histocompatibility barrier without inducing overt GvHD. The transplanted animals become tolerant to both donor and host grafts.

C57BL/6 mice were injected subcutaneously with Lewis-tumor cells and syngeneic spleen lymphocytes. When these lymphocytes had been preincubated with a thymic extract, tumor development was impaired: the tumor weight and volume were significantly decreased and the number and size of lung metastases were strikingly diminished. In addition, the increase in weight of adrenals and spleen, common in tumor-bearing animals, was not observed. Lymphocytes incubated with a thymic factor preparation can therefore enhance the defense against this tumor.

This paper reports three cases of lupus erythematosus (LE) associated with thymomas, all of which were discovered after the onset of LE. The diagnosis of LE was established by clinical and laboratory data. Histologically, the thymomas were of the three main types: epithelial, lympho-epithelial, and predominantly fusiform. In two cases, thymectomy did not modify the course of LE; in one vase, the absence of cytoplasmic secretory vesicles in the epithelial cells and the non-labelling of these cells by an antiserum thymic factor monoclonal antibody represented direct evidence of a functional thymus deficiency. Antikeratin antibodies were used to distinguish thymomas from lymphomas; epithelial thymomas exhibited the same keratin specificities as normal thymus, although the labelling pattern (net-like) was different. Labelling of epidermis with the same antiserum confirmed the theory of an epidermis-thymus relationship.

Homozygous sickle cell disease patients have an increased risk of developing severe infections, probably because of impaired immunity. Cellular immunity was studied in thirty-two patients with S homozygous haemoglobin (SS) and compared to 32 A homozygous haemoglobin (AA) healthy subjects. A leukocytosis was observed but with a significant diminution of T4 and T8 proportions in sickle cell patients. B lymphocytes, concanavalin A, phytohaemagglutinin, and phorbol myristate acetate-induced lymphocyte proliferation were not different between the two groups except for enhanced pokeweed mitogen stimulation in SS patients. In contrast, addition of autologous sera to mitogen-induced cultures resulted in a potentiation of lymphocyte proliferation significantly greater in patients with S homozygous haemoglobin when compared to subjects with A homozygous haemoglobin. This highly amplified mitogen-induced response of lymphocytes by SS autologous sera, when compared to AA autologous sera, was not observed when these sera were added to lymphocytes obtained from an allogenic healthy individual. In vivo interleukin 2 production and natural killer activity were not different between SS and AA individuals. We conclude that there are functional abnormalities of cell-mediated immunity in patients with sickle cell anaemia and the SS lymphocyte activation by autologous sera was probably due to infectious and drepanocytic antigenic determinants contained in SS serum.

Over 300 individuals have received fetal liver transplants for a spectrum of disorders including immunodeficiencies, aplastic anemia, leukemia and genetic disorders. In some instances, the objective has been to reconstitute the immune system from fetal liver-derived lymphoid stem cells. In aplastic anemia and leukemia two distinct approaches have been used: engraftment of fetal liver-derived hematopoietic stem cells or attempts to stimulate recovery of autologous hematopoiesis via factors produced by fetal liver. In genetic disorders, partial engraftment of fetal liver-derived hematopoietic and hepatic cells has been investigated. This report critically reviews data presented at a symposium on fetal liver transplantation in New-Delhi, 1-5 February 1986.