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This article summarizes the hematological and immunological consequences of splenectomy in children. The hematological consequences are mainly noticed in the number of white cells and platelets. Indeed, platelets can remain elevated several weeks and even several months after surgery. Rarely antiplatelets aggregant drugs will be given if the platelet count is higher than one million per cubic millimeter. The immunological consequences of splenectomy in children are mainly manifested if the surgery was performed before the age of 6 or 7. The major consequence is a rapid and often lethal infectious complication. In diseases like thalassemia major or lymphoma, there is an increased risk of infection. The immunological mechanism seems linked to a decreased antibody formation in presence of higher antibody titre needed by the reticuloendothelial system to eliminate the microorganisms responsible for the infection. Therapeutic measures can be proposed: delay the age of splenectomy, leave some splenic tissue, administration of penicillin and lastly immunization against some antigens mainly responsible of the infection.

The peripheral lymphocytes of a patient with prodromal acquired immune deficiency syndrome contained giant multivesicular bodies. These were specifically stained by immuno-gold labelled polyclonal antibodies against the major core protein p24 of bovine leukaemia virus and human T cell leukaemia virus I. Moreover, the patient's serum was positive for bovine leukaemia virus by the ELISA method.

Lynestrenol, a synthetic steroid progesterone-like substance, has been found to enhance the stimulation of normal human lymphocytes. It increases the phytohemagglutinin response and the mixed-lymphocyte cultures. This augmentation can, in some cases, be higher than 100% of the phytohemagglutinin or mixed lymphocyte reaction baseline. There is no significant effect on the pokeweed mitogen stimulation. It is concluded that lynestrenol can modulate, "in vitro", some T cell functions.

In vitro effects of three immunosuppressive drugs, cyclosporin, methylprednisolone, and 6-mercaptopurine (the active moiety of azathioprine), have been assessed by various proliferative assays including mixed lymphocyte reaction (MLR), concanavalin A (Con A), OKT3 monoclonal antibody (MoAb), phorbol myristate acetate (PMA), and A23187 ionophore-induced mitogenesis. The influence of these drugs on the capacity to produce interleukin-2 (IL-2) after phytohemagglutinin (PHA) induction has also been evaluated. Depending upon the nature of the stimulus used, differences were found among the three immunosuppressive drugs, suggesting that different T-cell activation mechanisms are involved. The response to alloantigens (MLR) or to mitogens (Con A and OKT3 MoAb) resulted in significant inhibition by all three immunosuppressive agents, although 6-mercaptopurine was somewhat less potent at the pharmacological concentration of 0.1 microgram/ml. When PMA (which activates cells through protein kinase C) and A23187 ionophore (which enhances Ca2+ influx into the cytoplasm through membrane channels) were used, dissociations were observed, suggesting that drugs act differentially on early stages of T-cell activation. Inhibition of PMA-induced activation by cyclosporin was very limited in contrast to the potent effect of methylprednisolone. The latter drug could thus preferentially affect an activation step involving the PMA receptor. Furthermore, the inhibitory influence of methylprednisolone on A23187 ionophore-induced proliferation was equivalent to that of cyclosporin, while 6-mercaptopurine had no effect except at the high concentration of 1 microgram/ml in which direct inhibition of DNA replication is probably involved. IL-2 production induced by PHA was strongly blocked by methylprednisolone and cyclosporin but not by 6-mercaptopurine, at least when a pharmacological concentration was used. In view of the relative independence of ionophore-induced activation from the influence of IL-2, this response could be linked to the intrinsic mechanism of antiproliferative activity of immunosuppressive drugs which probably act mostly at the level of IL-2 production. These results also provide a cellular approach to the synergies between immunosuppressive drugs. Most of the tests are more strongly affected by cyclosporin than by 6-mercaptopurine, probably explaining the potent steroid-sparing effect of cyclosporin.

The effect of vinblastine and colchicine on human blood T cell rosette formation with sheep erythrocytes was assessed. For some concentrations, both drugs increased the percentage of active T rosettes. In contrast, the total T rosette percentage was decreased by the same drugs. These experiments further confirm the heterogeneity of T cell rosettes and suggest the existence of three types of sheep red blood cell receptors on human blood T lymphocytes.

High levels of tumor necrosis factor-alpha, interleukin-2, and gamma-interferon appeared in the circulation of kidney transplant recipients after the first injection of the monoclonal antibody OKT3. This initial injection was systematically followed by fever. The three cytokines were released in all patients (n = 9), with peak serum levels of tumor necrosis factor occurring 1 hr after OKT3 injection and those of interleukin-2 and gamma-interferon after 2 hr. Cytokines were not released after the second and third OKT3 injections, when CD3+ cells had disappeared from blood. These findings suggest that circulating cytokines are released by T cells after activation by OKT3. These cytokines are probably involved in the systematic reactions observed after injection of OKT3.

Immune-mediated mechanisms and genetic factors are believed to be involved in the pathogenesis of Crohn's disease. We studied T- and B-cell subpopulation proportions and various functional assays, including proliferative responses to PHA and Con A, Con A-induced suppressive activity, and natural killer cell assay toward the K562 cell line, in the peripheral blood of 22 patients with inactive familial Crohn's disease and their 35 healthy relatives including nine families. HLA-A, -B, and -DR antigens were determined in all the subjects. With the exception of minor abnormalities of suppressor cell activity present in some relatives of two families, neither significant impairments of immunological parameters in patients or their relatives nor concordant segregation of HLA haplotypes and disease were observed. These data indicate that peripheral immune abnormalities previously described in patients with Crohn's disease do not constitute primary factors involved in the disease itself and that familial incidence in Crohn's disease cannot be linked to immunological markers presently studied.

We report a case of a double M component appearing in the course of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD). Lymphocyte function, especially the decrease in T-suppressor cells, could play an essential role in the mechanisms of the disease. In the presence of an M component, the prognosis for AILD seems to be more pessimistic. Arguments for considering AILD as a premalignant disease are reviewed.

A new approach to rapidly determine human mixed-lymphocyte culture (MLC) is presented. It is based on the identification of an early increase in the percentages of active T rosettes (TEa) in the MLC tubes. Active T rosettes represent a subset of T cells with high-avidity receptors for sheep red blood cells. All subjects tested had a full determination of their HLA-A, -B,-C, and -D-DR. The following observations were made: (1) When the MLC was done between 10 pairs of fully HLA-A,-B,-C, and -D-DR incompatible subjects, a highly significant increase (P less than 0.001) was observed after 2 hr of MLC in all pairs tested. (2) The same increase was observed in all 10 pairs of identical HLA-A,-B, and -C subjects but not sharing HLA-D-DR antigens. (3) In the cases of HLA-D-DR semi-identity, the increase in TEa percentages was less marked after 2 hr of culture (P less than 0.05) and delayed in time to reach the significance level of P less than 0.001 after 4 hr of culture. (4) In contrast, in the case of full HLA-D-DR identity, no increase in TEa percentages was observed at any time in the culture (2, 4, and 24 hr and 4 days) in nine pairs of subjects tested (two pairs of identical twins). In conclusion, these results indicate that the active T rosette MLC is a rapid approach (2 hr) to evaluate HLA-D-DR differences without influence by HLA-A,-B, or -C. As such it is an early marker for HLA-D-DR determination and may become a useful tool in the field of human histocompatibility to study the HLA-D-DR region.

The influence of inosiplex upon various in vitro leucocyte assays was studied in normal individuals. It was found that the drug increases the response of bidirectional and unidirectional mixed lymphocyte cultures at concentrations of 200, 300, and 500 microgram/ml. It also significantly increases the percentage of active T rosettes (concentration range: 50 to 500 microgram/ml) and the percentage of autologous red cell T rosettes (concentration: 100 microgram/ml). In contrast, inosiplex did not modify the percentage of total T rosettes and EAC rosettes. Inosiplex increases the number of nonadherent leucocytes in the leucocyte adherence inhibition test at a concentration range between 100 and 300 microgram/ml. Finally, inosiplex also increases the percentage of monocytes phagocytizing yeast at a concentration between 50 and 500 microgram/ml. These data indicate that inosiplex enhances the function of normal human T cells, monocytes, and possibly neutrophils. Therefore inosiplex appears to have immunostimulant properties.

The interferons are known to induce the expression of cell surface determinants. Gamma interferon, in particular, has been shown to enhance class II antigens (DR) on the cell surface. We used this gamma interferon to induce beta 2-microglobulin (beta 2 mu) and a minor, sex related transplantation antigen called H-Y on the surface of B lymphoma cells. The antitumor effect of interferon could thus be at least twofold: an antiviral effect already known and an increase of intercellular recognition (by cell surface marker enhancement); allowing the tumor cells to be "seen" better by cells of the immune system.

The clinical and cytogenetic findings of a patient with a preleukemic state and trisomy 11 are reported. Trisomy 11 was present as the sole karyotypic alteration at the time of overt leukemia. Trisomy 11 presents an additional chromosomal abnormality not previously described in preleukemia.

Renal transplant recipients receiving low maintenance immunosuppression (azathioprine, 2 mg/kg/day, and prednisolone, 10 mg/day), tolerating their transplants well, and without viral infection disclose a profound depression of NK activity as assessed by 51Cr-release assay. By combining the analysis of the different steps of cytolysis with the agarose single-effector assays and the estimation of circulating large granular lymphocytes (LGL), the defect is shown to be due to a significant decrease of the number of NK cells capable of binding (% target-binding cells 2.0 +/- 0.3 versus 5.7 +/- 0.7 in normals, P less than 0.001) and killing (% cytotoxic target-binding cells 12.4 +/- 1.9 versus 22.0 +/- 0.5 in normals, P less than 0.001) of targets. There is also a significant reduction (P less than 0.001) of both percentages (1.0 +/- 0.2 versus 3.3 +/- 0.4 in normals) and absolute values (9.8 +/- 2.4 versus 62.3 +/- 8.0/microliters in normals) of LGL. These observations indicate that depressed NK activity is due mostly to depletion of NK cells. Functional impairment of NK cells can also be involved. Lack of direct in vitro effects of drugs (6-mercaptopurine, hydrocortisone, and methylprednisolone) at concentrations likely to be reached in vivo during treatments and relative resistance of NK activity after in vivo steroid administration suggest that immunosuppressive drugs act at the precursor cell level or on regulatory mechanisms. Despite functional integrity of two suppressor cell systems of allogeneic NK activity (suppression induced by preculture of lymphocytes with Con A and suppressor granulocytes) in immunosuppressed patients, tested on normal NK cells, NK cells of immunosuppressed patients did not disclose greater susceptibility to Con A-induced suppression. This analysis indicates that the depletion phenomenon is probably a major mechanism in NK depression of patients receiving immunosuppressive drugs.

A fluorometric assay is described for the detection of red blood cell antibodies. The assay reveals as little as 600 molecules of bound, fluoresceinated rabbit anti-human IgG antibodies per erythrocyte. Eleven patients with possible autoimmune erythrocyte disorder and negative direct antiglobulin test were studied by the fluorometric assay. The outcome of the fluorometric assay was compared with that of the human allogeneic rosette test. Results obtained by the two methods were in complete agreement. Five of the patients were shown to possess unexpectedly high levels of erythrocyte-bound IgG in spite of a negative, direct antiglobulin test. These findings and the validity of the fluorometric assay are discussed.

This paper summarizes the data related to the relations between enkephalins and endorphins and the immune system. It is shown that lymphocytes have surface receptors for endorphins and enkephalins. Furthermore, endorphins and enkephalins can influence several immune functions such as antibody synthesis, lymphocyte proliferation, and natural killer cytotoxicity. It is thus possible that the receptors play a functional role. In view of these observations, several hypotheses are proposed. Endorphins and enkephalins are physiological regulators of the immune response (two-hit opioid peptide lymphocyte receptor hypothesis) and they are humoral mediators between the central nervous system and the immune system. They may play a pathogenic role in a variety of diseases with primary or secondary immunological defects. Finally, enkephalins and endorphins can be considered as immunomodulators and modifiers of the biological response and as such may become a tool in the field of immunotherapy.

Peripheral leucocytes from healthy blood-donors were preincubated in polyarthritic serum or normal serum and their adherence to glass tubes in the presence of synovial extract of control extract was measured. Serum from patients with inflammatory polyarthritis significantly reduced the proportion of leucocytes which adhered in the presence of synovial extract, but not control extract. Serum from patients with polyarthritis with visceral manifestations, patients with degenerative or metabolic polyarthritis or rheumatic fever, and controls did not have this effect. The leucocyte sensitising activity was eluted in the IgG fraction of serum, and could be absorbed by both synovial extract and IgG. It is thus probably IgG rheumatoid factor. In vivo, armed leucocytes in polyarthritic patients may bind to synovial tissue initiating or perpetuating the articular inflammatory process.

Lynestrenol, a synthetic progesteronelike drug, was assessed for its influence upon human leukocyte tests in vitro and in vivo. Lynestrenol markedly enhanced the lymphocyte response to phytohemagglutinin, the mixed lymphocyte culture, the active T-rosette test, the autologous red cell rosettes, and the number of nonadherent cells in the leukocyte adherence inhibition test. All these results indicate a stimulatory effect upon human T-cell function. Lynestrenol also increased monocyte phagocytosis. Lynestrenol delayed the appearance of malignant tumors in hamsters. Finally, lynestrenol, given to lung cancer patients, increased their T-cell functions as evaluated by skin tests. T-cell rosettes, and phytohemagglutinin response. Lynestrenol does possess immunostimulatory properties.

Monoclonal antibodies specific to human T lymphocyte receptors are currently being used to define the biochemical structure of these proteins as well as of functionally distinct cell subsets. Since one of the antibodies (OKT3) recognizing the T3 (CD3) receptor mimics vital physiological processes involved in the activation of the immune system and has been successfully used as a therapeutical agent, we investigated one of the mechanisms underlying this antibody-receptor interaction. Our results show that after binding of OKT3, the complex (OKT3-T3) disappears rapidly from the cell surface. Using electron microscopy, we found that this down-regulation is due to the internalization of the complex. Parallel experiments performed on the T11 (CD2) and T4 (CD4)/AIDS retrovirus receptor indicate that the same mechanism applies for the down-regulation of those molecules. These data suggest that the T3, T11 and T4 receptors have a behavior comparable to other well characterized, hormonal and viral receptors; they provide information on the metabolization pathway of surface receptors and on the possible intracellular penetration of ligands like the HTLV-III/LAV agent in human T lymphocytes.

Human red blood cells can bind to human peripheral blood lymphocytes. The percentage of these rosette-forming cells (TEh) depends on various conditions including temperature and the use of selected foetal calf serum. It is improved by toluidine blue. Prior incubation at 37 degrees C is not necessary. ABO and rhesus antigen D have no influence on rosette formation. Analysis of human rosettes in T and B cell rich populations, in patients with chronic lymphatic leukaemia or T cell lymphoma provides further evidence that human autologous and allogeneic rosette-forming cells belong to a T cell subpopulation.

A woman with a T cell lymphoproliferative malignacy and heterozhgosity at the X chromosome-linked locus for glucose-6-phosphate dehydrogenase (G-6PD) isoenzymes was studied to find the clonal origin of her circulating neoplastic T cells. The red blood cells, polymorphonuclear cells, whole mononuclear cells, and T cell-depleted mononuclear cells contained both A and B isoenzymes of G-6-PD. In contrast, the tumor cells, separated by using their capacity to form rosettes with sheep red blood cells, contained only the B isoenzyme of G-6-PD. This observation strongly suggests the monoclonality of this T cell malignancy.

We report the case of an African patient with acquired immune deficiency syndrome (AIDS) who had a chronic cryptococcal rectal abscess with anal fistula and a disseminated neurologic and bony fungal disease, associated with pulmonary infiltration due to Pneumocystis carinii. The anal lesion was surgically excised because of failure of the medical treatment. Although clinical intestinal cryptococcal involvement is quite rare, the experience here recorded should draw physicians' attention to the possibility of an insidious disseminated disease in AIDS patients.

This paper introduces an immunogold silver staining procedure for the identification of human lymphocyte subpopulations in optical microscopy. The described procedure is performed on cells in suspension (peripheral mononuclear cells or blood buffy-coat cells) and uses conventional antilymphocytic monoclonal antibodies (OKT3, OKT4, OKT8, OKIa1) followed by gold-labeled secondary antibodies (GAM G30 or G40) and silver sensitization. It ends with the preparation of permanent records (smears or cytocentrifuge preparations) which are counterstained with standard panoptic Wright or May-Grünwald Giemsa stain. In all cases, the surface antigens appear as numerous black dots on the lymphocytes, with a strong labeling reaction that allows one to clearly distinguish between negative and positive cells. The comparison with immunofluorescence microscopy in normal individuals and in patients indicates that this new immunostaining technic is specific and more sensitive. It has the advantages of using small amounts of blood (1 mL per test), of being rapid (three hours), and of allowing the simultaneous immunophenotypic and morphologic evaluation on smears. This makes it suitable for the analysis of other biologic specimens not already accessible with immunofluorescence. The labeled preparations apparently can be stored indefinitely, and are thus useful for longitudinal studies in the same patient. Finally, the method does not require sophisticated equipment and may be subjected to automated analysis.

Human recombinant interferon-alpha 2C was given to 4 patients with essential thrombocythaemia. Three patients achieved a complete remission on day 19 +/- 1 and the 4th achieved a partial remission. After 4 weeks of induction therapy, a maintenance therapy of twice weekly intramuscular injections of 5 X 10(6) or 10 X 10(6) IU was instituted but did not maintain the remission.

The action of various concentrations of levamisole on normal human lymphocytes was investigated in vitro. The action of levamisole (10(-1) mg-10(-8) mg%) upon rosette formation was studied using two rosette assays with SRBC (the active and the total T-rosette tests) to quantify T cells and the EAC assay to quantify B cells. Levamisole at only 10(-3) mg% significantly increased the percentage of both active and total T cells. In sharp contrast, levamisole between 10(-2) and 10(-7) mg% significantly decreased the EAC percentage. It is concluded that levamisole promotes a better expressivity of T-cell receptor and decreases the expressivity of C3 receptor.

18 patients with osteogenic sarcoma were followed by serial measurements in vitro of tumor-specific cell-mediated cytotoxicity and of "active" and total rosette-forming T-cells. 13 of these patients have had or are currently receiving injections of osteogenic sarcoma-specific dialyzable transfer factor derived from healthy donors. In three patients with very small lesions, cytotoxicity was high before amputation and decreased within 2 mo after removal of tumor. Cytotoxicity was low at time of diagnosis in all patients with large tumor masses. The cytotoxicity of the patients' lymphocytes increased after administration of tumor-specific transfer factor in all patients so treated. Patients receiving nonspecific transfer factor showed evidence of declining cell-mediated cytotoxicity. Tumor-specific transfer factor may produce an increase in cell-mediated cytotoxicity to the tumor in patients with osteogenic sarcoma. This possibility is suggested by the pain and edema that occurred in the area of the tumor in patients who had metastatic disease when therapy was started and by lymphocytic infiltrates in the tumor, as well as by the increase in cell-mediated cytotoxicity and the increase in percentage of active rosette-forming cells from subnormal to normal. Serial measurements of cell-mediated cytotoxicity are helpful in monitoring the efficacy of transfer factor and other modes of therapy in these patients, and these measurements are the best available criteria for selection of donors of tumor-specific transfer factor.

Peripheral blood mononuclear cells from 15 patients with alcoholic cirrhosis (AC) and 15 matched healthy controls were tested for their proliferative response to mitogens such as PHA and con A, tumour promoter PMA and OKT3 monoclonal antibody, for their capacity to produce IL-2 and to respond to recombinant IL-2 (rIL-2). The expression of IL-2 receptor (Tac) together with two other activation markers, the receptor for transferrin (T9) and Ia antigen have also been assessed. A profound decrease of proliferative response was observed after stimulation by lectins and PMA. IL-2 production was significantly decreased (0.39 +/- 0.07 versus 0.82 +/- 0.009 units; P less than 0.001) in alcoholic cirrhosis. Resting PBMC of these patients disclosed spontaneous responsiveness to rIL-2 without requiring prestimulation with PHA as observed in healthy subjects. This abnormal response was associated with significantly increased percentages of Tac (19.4 +/- 3.1 versus 5.1 +/- 1.1%; P less than 0.001), T9 (13.6 +/- 3.3 versus 6.6 +/- 1.1%; P = 0.04) and Ia positive cells (21.4 +/- 5.4 versus 11.5 +/- 1.4; P less than 0.05) in alcoholic cirrhosis. Defective proliferative activity and impaired IL-2 production, combined with an increase of lymphocytes bearing T cell activation markers and of rIL-2 responsiveness could represent in vitro correlates for mechanisms leading to immunoregulatory disturbances in cirrhotic patients.