OKT8 Cells Research Articles

Bone marrow transplantation for poor prognosis ALL in children. Relapse rate, infections and immunological recovery.

SummaryBetween 1980 and 1985, 11 children (8 boys, 3 girls aged between 18 months and 16 years) with poor prognosis ALL (Acute Lymphoblastic Leukaemia) were treated with ablative therapy and allogeneic bone marrow transplantation. Six patients (55 %) are alive at the time of evaluation , one died in aplasia, two died of interstitial pneumonia and two died of bone marrow relapse.The median time to recover a neutrophil count of 0.5 x 109/1 was 19 days. The total number of lymphocytes reached normal values after 6 months. The percentage of OKT3 cells (mature T lymphocytes) was slightly depressed during the first 3 months. For at least 9 months, the number of OKT4 (helper lymphocytes) was low with a normal number of OKT8 (suppressor lymphocytes) resulting in an OKT4/OKT8 inverted ratio.As far as B lymphocytes function was concerned, there was a moderate decrease of the immunoglobulin levels. Normal values were obtained after 3 months for IgG, 6 months for IgA and IgM.All the immunological parameters tested returned to normal within one year after the bone marrow transplantation.

Risk of transmission of human immunodeficiency virus (HIV) by heat-treated factor VIII concentrates in patients with severe hemophilia A.

At the change from unheated to heat-treated Factor VIII concentrates for the treatment of hemophilia A, 17 severe adult hemophiliacs (mean monthly dose, 4927 IU) were evaluated prospectively for signs of infection with human immunodeficiency virus (HIV). Viral serology and lymphocyte subpopulations (OKT3, OKT4, and OKT8-positive cells) were examined monthly for 1 year. One patient seroconverted for HIV in the enzyme-linked immunoabsorbent assay but was positive on the Western blot analysis from the outset. There was a slight but significant increase in OKT4+ cells and OKT4/OKT8 ratio. These data suggest that heat-treated Factor VIII concentrates even when used in large amounts have a low risk of transmitting HIV.

Factors influencing circulating OKT8 cell phenotypes in patients with multiple sclerosis.

Peripheral blood OKT8 cell phenotypes were correlated with measurements of plasma cortisol and serological evidence for exposure to 15 infectious agents, in longitudinal studies involving 13 patients with multiple sclerosis, 13 of their siblings, nine spouses and 13 unrelated controls; 44/48 individuals were HLA typed. Neither circadian rhythms, nor exposure to any one infectious agent accounted for the serial changes in OKT8 cells but there was an association between the presence of HLA-DR2 and periodic reductions in OKT8 cells irrespective of clinical status. Taken with previously reported serial observations in patients and cohabiting relatives, this finding provides indirect evidence for an interplay between environmental and genetic factors in determining OKT8 cell phenotypes in multiple sclerosis.

Presence of in vivo-activated T-cells expressing HLA-DR molecules and IL-2 receptors in peripheral blood of patients with nasopharyngeal carcinoma.

Epstein-Barr Virus (EBV) is associated with two malignant diseases, African Burkitt's Lymphoma (BL) and Undifferentiated Nasopharyngeal Carcinoma (UNPC). North Africa is a geographical area with a high incidence of NPC. Our purpose in this study was to explore cell-mediated immunity of peripheral blood lymphocytes (PBL) from patients with UNPC and DNPC. We found an elevated percentage of OKT8 cells and of large granular lymphocytes (LGL) (30-35% HNK-I-positive cells) compared to PBL from healthy matched individuals. PBL from NPC patients contained 35% HLA-DR-positive and 30% Interleukin-2 (IL-2) receptor-positive circulating lymphocytes. PBL from NPC patients exhibited a normal proliferative response to phytohemagglutinin (PHA) and Concanavalin A (Con A) and an increased response to pokeweed mitogen (PWM). Natural killer (NK) activity towards K562 cells was low in our patients who, in addition, exhibited no lytic activity against HLA-matched EBV-transformed B cells. This lack of cytotoxicity against an EBV-transformed B-cell line cannot be explained by an impairment of IL-2 secretion, and is probably a result of the presence of high numbers of OKT8 suppressor T cells.

Peripheral blood and tumor-infiltrating mononuclear cell analysis in esophagus and head and neck cancer.

We show a significantly decreased number of OKT3+ lymphocytes in the peripheral blood (PB) of cancer patients mainly due to a reduced number of OKT4 cells. OKT8 cells were also somewhat reduced. The numbers of DR+ and interleukin-2 receptor-bearing T-cells were significantly increased in patients. The tumor-infiltrating cells included OKT4+ AND OKT8+ lymphocytes. There was a significant correlation between the proportion of activated T-cells in PB and in tumor, as shown by DR and interleukin-2 receptor expression.

Dynamic Relationship between Hydrolytic Enzymes and the Immune System in Rheumatic Diseases

We performed longitudinal studies on patients with two different types of rheumatic diseases, systemic erythematodes (SLE)-like and rheumatoid arthritis (RA)-like, to elucidate the relationship between hydrolytic enzymes and immunological disturbances. The plasma levels of 9 hydrolytic enzymes and subpopulations of T-cells, OKT4 and OKT8 cells, in peripheral blood were checked periodically for 6 months. Analysis of the obtained data by a method of spectral analysis utilizing Akaike's autoregressive approach clearly demonstrated the existence of a dynamic network between the hydrolytic enzymes and the immune system. In testing the behavior of this network, we obtained data suggesting that the response of OKT4 cells when an impulse is given to OKT8, is abnormal in the SLE-like syndrome, and that this abnormality may be related to the malfunction of suppresser T-cells in this pathological condition. Principal component analysis suggested that the behaviors of kallikrein and carboxypeptidase B are quite different between the two types of immunological disturbances. The network relationships between the hydrolytic enzymes and the immune system seem to play important roles in the pathophysiology of rheumatic diseases.

Abnormality of alveolar lymphocytes in sarcoidosis

Brochoalveolar lavage (BAL) were performed in 28 patients with sarcoidosis and in 31 normal subjects. In control subjects, the numbers of mononuclear cells and macrophages in smokers were significantly increased in comparison to nonsmokers (p<0.001 in each cases), but the numbers of lymphocytes were not.In both smoking and nonsmoking untreated sarcoid patients, the proportion and number of lymphocytes were significantly increased compared to corresponding controls (p<0.05, p<0.001). The number of lymphocytes in patients not receiving prednisolone were not significantly increased than patients who were receiving prednisolone. No difference in numbers of lymphocytes was found between smoker and nonsmoker patients. BAL lymphocytes were identified as T lymphocytes (88.9±6.2%). Furthermore, untreated patients had significantly higher percentages of OKT-4 (+) T helper cells and significantly lower OKT8 (+) T suppresser cells within the alveolar lymphocytes population than that of normal nonsmokers (p<0.01 in each cases). Alveolar lymphocytosis is significantly correlated with the activity of lymphocyte blastogenesis induced by Propionibacterium acnes (p<0.01), and the ratio of OKT-4 (+) cells to OKT-8 (+) cells correlated with the activity of serum angiotensin-converting enzyme (p<0.05). However, neither the radiological stage nor the degree of Gallium-67 uptake in lung parenchyma was related to the lymphocyte number and the ratio of OKT-4/OKT-8 in BAL fluids.The lymphocytosis of the bronchoalveolar space indicates the disease activity at the site of pulmonary space, and Propionibacterium acnes may play some role in the induction of the pulmonary lymphocytosis in sarcoidosis patients.

Healthy HTLV-I carriers in Japan: the haematological and immunological characteristics.

The haematological and immunological characteristics of 34 healthy anti-HTLV-I antibody-positive individuals (HTLV-I carriers) in southwestern Japan were examined. No significant difference was noted between carriers and the controls in counts of RBC, WBC and the absolute number of lymphocytes. The serum IgG in the carriers was higher than that of the controls. The percentages of OKT4, OKT8, OKIa1 and B1-positive cells were found to be normal in the peripheral blood of the carriers, whereas the percentages of OKT11 and anti-Tac-positive cells were significantly higher in the carriers than in the controls. A correlation was observed between the percentages of anti-Tac-positive cells and the titres of anti-HTLV-I antibody in the carriers. After a 72 h incubation of peripheral blood lymphocytes with medium alone, the percentage of anti-Tac-positive cells tended to decrease in the controls, but to increase in carriers, with the appearance of large blastoid cells resembling blastic transformed lymphocytes cultured with mitogen. Tac and Ia antigens were markedly expressed on these large blastoid cells.

Characterization of human cytotoxic lymphocytes directed against cells infected with typhus group rickettsiae: evidence for lymphokine activation of effectors.

An in vitro culture and assay system was used to determine whether cytotoxic lymphocytes are generated in humans after rickettsial infection. Peripheral blood mononuclear cells (PBMC) were obtained from six individuals with serologic evidence of prior infection with typhus group rickettsiae and from six nonimmune individuals. After PBMC from immune individuals were stimulated in vitro for 7 days with rickettsial antigen, they were capable of lysing typhus group rickettsia-infected, autologous phytohemagglutinin (PHA)-induced blasts, but not uninfected PHA-blasts. No cytotoxic effector cells were generated when either PBMC from immune individuals were placed in culture for 7 days without antigenic stimulation, or when PBMC from nonimmune individuals were stimulated in vitro with antigen for 7 days. Freshly isolated PBMC from immune donors were also unable to lyse typhus group rickettsia-infected autologous PHA-blasts or an autologous rickettsia-infected lymphoblastoid cell line (LCL). Neither supernatants from antigen-stimulated cultures of PBMC from immune donors nor recombinant human interferon-gamma were capable of significantly lysing typhus group rickettsia-infected PHA blasts by this assay. Populations of cytotoxic effector cells depleted of OKT3, OKT4, or OKT8-positive cells by treatment with the respective monoclonal antibodies and complement were assayed for their cytotoxic capacity. The results suggest that the cytotoxic effector cell population is predominantly OKT3 and OKT8-positive, but OKT4-negative. Positive selection with the use of a fluorescence-activated cell sorter also suggested that most of the cytotoxic effector cells are OKT8-positive. PBMC from immune donors after in vitro stimulation with rickettsial antigen were capable of significantly lysing infected autologous LCL or infected HLA-mismatched LCL as compared with the respective uninfected controls. In addition, PBMC from either immune donors or nonimmune donors after stimulation in vitro for 7 days with media containing purified lymphokines were capable of significantly lysing autologous infected LCL as compared with the uninfected autologous control. We conclude that lysis of cells infected with typhus group rickettsiae is mediated by a lymphokine-activated killer.

Selective partial IgM deficiency: functional assessment of T and B lymphocytes in vitro.

Seven patients with selective IgM deficiency (SIgMD) were studied for cell surface immunoglobulin (SmIg), T-cell subpopulations, and immunoglobulin (Ig) synthesis in vitro by peripheral blood lymphocytes (PBL). Serum IgM levels were less than 25 mg/dl, while IgA, IgG, and IgD were within normal levels. The patients had respiratory or urinary tract infections and two were diagnosed as having systemic lupus erythematosus (SLE). T/B-cell ratios in PBL were within normal ranges. Percentage ratios of B cells bearing SmIg were normal in five patients and decreased in two; however, normal values were seen after 7 days of culture in the presence of PWM. OKT4/OKT8 ratios decreased in five of seven patients, in whom two were due to a decrease in OKT4 and two to an increase in OKT8 cells. One showed a decrease in OKT4 and an increase in OKT8. Analysis of lymphocyte function for Ig synthesis in vitro, using a coculture of counterpart T and B cells from healthy individuals and patients with SIgMD, revealed that the increased function of IgM isospecific suppressor T cells (Ts) was responsible for the IgM deficiency in all seven patients.

Circulating T- and B-Cell Abnormalities in Cutaneous Lupus Erythematosus

Seven patients with subacute cutaneous lupus erythematosus (SCLE), 11 patients with discoid lupus erythematosus (DLE), and 17 age-, sex-, and race-matched controls were examined for the number of circulating peripheral blood T and B cells, immunoglobulin (Ig) synthesizing cells, and Ig secreting cells. A significant alteration in T-cell subsets was detected only in SCLE patients who manifested a decreased number of OKT8 cells. Circulating B cells were significantly increased only in the SCLE patients. The percentage of Ig synthesizing and secreting cells was significantly increased in both the DLE and the SCLE groups. The somewhat unexpected increase in Ig synthesizing and secreting cells in the DLE patients could not be accounted for by antimalarial treatment or the concurrence of the HLA-DR3 phenotype. There was, however, a weak but significant correlation between the degree of B-cell activation in the DLE patients and the number of American Rheumatism Association criteria for the classification of systemic lupus erythematosus (SLE) possessed by these patients. These data demonstrate that abnormalities in B-cell function similar to those seen in SLE can also be found in a group of lupus erythematosus patients whose clinical disease expression is limited predominantly to the skin.

Immunoregulation of B lymphocyte colony formation by T cell subsets in patients with chronic lymphocytic leukemia

Normal B lymphocytes are activated, proliferate, and then differentiate into plasma cells and secrete immunoglobulin (Ig). We have reported that chronic lymphocytic leukemia (CLL) T4 cells help and CLL T8 cells lack suppressor effects on Ig synthesis by normal B cells (Blood 62:767, 1983). We have now explored the earlier phase, proliferation, using B cell colony formation; in semisolid media. B lymphocyte colonies from normal individuals and from patients with CLL were grown in 0.3% agarose overlayed with T cells or T cell subsets and the B cell mitogen staphylococcal protein A. Enriched T cells, OKT4 or OKT8, were obtained either by sheep erythrocyte rosettes or depletion of OKT8 or OKT4 cells by monoclonal antibody or complement, respectively. Twenty thousand B cells from normal subjects yielded 65 +/- 9, 64 +/- 7, and 19 +/- 6 colonies with autologous unfractionated T-, OKT4-, or OKT8- positive cells, respectively. This compared to 29 +/- 11, 81 +/- 11, and 15 +/- 4 colonies from patients with CLL with added autologous unfractionated T-, OKT4-, or OKT8-positive cells. To determine whether the fewer number of colonies in both normal subjects and patients with CLL with OKT8-positive cells was due to suppression or lack of help, the number of OKT4-positive cells was held constant, and OKT8-positive cells were added in increasing numbers. No suppression of colony formation could be demonstrated. Furthermore, the addition of increasing numbers of concanavalin A (Con A)-activated OKT8-positive cells did not suppress colony formation. These results suggest that the CLL T cell subsets behave in a functionally similar manner to normal T cell subsets, namely, (1) that normal and CLL B cell colony growth is helped by OKT4 cells; and (2) in contrast to immunoglobulin secretion by B cells, neither normal nor CLL OKT8 cells, unstimulated or activated by Con A, suppress B cell colony growth.

Immunoregulation of B Lymphocyte Colony Formation by T Cell Subsets in Patients With Chronic Lymphocytic Leukemia

Normal B lymphocytes are activated, proliferate, and then differentiate into plasma cells and secrete immunoglobulin (Ig). We have reported that chronic lymphocytic leukemia (CLL) T4 cells help and CLL T8 cells lack suppressor effects on Ig synthesis by normal B cells (Blood 62:767, 1983). We have now explored the earlier phase, proliferation, using B cell colony formation; in semisolid media. B lymphocyte colonies from normal individuals and from patients with CLL were grown in 0.3% agarose overlayed with T cells or T cell subsets and the B cell mitogen staphylococcal protein A. Enriched T cells, OKT4 or OKT8, were obtained either by sheep erythrocyte rosettes or depletion of OKT8 or OKT4 cells by monoclonal antibody or complement, respectively. Twenty thousand B cells from normal subjects yielded 65 +/- 9, 64 +/- 7, and 19 +/- 6 colonies with autologous unfractionated T-, OKT4-, or OKT8-positive cells, respectively. This compared to 29 +/- 11, 81 +/- 11, and 15 +/- 4 colonies from patients with CLL with added autologous unfractionated T-, OKT4-, or OKT8-positive cells. To determine whether the fewer number of colonies in both normal subjects and patients with CLL with OKT8-positive cells was due to suppression or lack of help, the number of OKT4-positive cells was held constant, and OKT8-positive cells were added in increasing numbers. No suppression of colony formation could be demonstrated. Furthermore, the addition of increasing numbers of concanavalin A (Con A)-activated OKT8-positive cells did not suppress colony formation. These results suggest that the CLL T cell subsets behave in a functionally similar manner to normal T cell subsets, namely, (1) that normal and CLL B cell colony growth is helped by OKT4 cells; and (2) in contrast to immunoglobulin secretion by B cells, neither normal nor CLL OKT8 cells, unstimulated or activated by Con A, suppress B cell colony growth.

Phenotypic modulation of T-lymphoblastic leukaemic cells with phorbol ester and leucocyte conditioned medium

Human T-lymphoblastic leukaemic cell line Karpas-45 (K-45) can be induced by phorbol 12-myristate 13-acetate (PMA) to become phenotypically more mature. Marker studies show that the percentages of E-rosette-positive (E +) and UCHT1 + cells are increased after exposure to PMA. Fluorescence of UCHT4 + and 2D1 + cells is increased although their percentages remain almost unchanged. OKT4 + and peanut agglutinin (PNA)-positive cells are greatly reduced. Terminal deoxynucleotidyl transferase (TdT) becomes negative. PMA treatment of K-45 cells after deletion of UCHT4 + cells suggest that marker changes detected by UCHT2, OKT4 and sheep red blood cells (SRBC) occur mainly in the UCHT4 + cells. The proliferation of PMA-treated K-45 cells is dramatically inhibited and the cell size becomes smaller. These results indicate that K-45 cells are induced to differentiate phenotypically towards a suppressor/cytotoxic T cell. However functional maturation can not be demonstrated. Phytohemagglutinin-stimulated leucoycte conditioned medium (LCM) is able to suppress DNA synthesis of K-45 cells and reduce the PNA + proportion. It seems that LCM can induce a limited degree of differentiation of K-45 cells.

Study on lymphocyte subsets and NK cell function in patients with dilated cardiomyopathy.

An acquired defect or damage of a subpopulation of suppressor T lymphocytes is reported in connection with autoimmune diseases. In the present study, the role of immunity was examined in 7 patients with dilated cardiomyopathy (DCM). The frequency of lymphocyte subsets using monoclonal antibodies and natural killer (NK) cell activity was evaluated to determine whether DCM patients had lymphocyte abnormalities that would support the hypothesis that the pathological mechanism of DCM is an immune disturbance. The peripheral lymphocyte counts were significantly lower in patients with DCM and higher in patients with ischemic heart disease (IHD) than in normal controls (NC) (p less than 0.01). The percentage of T cells, B cells, OKT4 and OKT8 positive cells was not statistically different among the three groups studied here, whereas the percentage of T gamma cells was significantly reduced in DCM patients (p less than 0.05). NK cell functional activity as tested in DCM and IHD patients was frequently deficient (22.1 +/- 19.3% in DCM, 13.8 +/- 3.0% in IHD, 37.4 +/- 12.7% in NC). Our results suggest that an imbalance in cellular immune reactions partly explain the pathogenesis of DCM.

T-cell subsets and natural killer activity in Plasmodium falciparum-infected children

Thirty children acutely infected by Plasmodium falciparum and suffering either benign uncomplicated malaria (17 cases), or cerebral malaria (13 cases), were investigated for T-cell number and subset distribution among peripheral blood mononuclear cells using OKT3, OKT4, and OKT8 monoclonal antibodies, and for natural killer (NK) activity using K562 cells as targets. They were compared to a group of 16 age- and sex-matched healthy Senegalese children. OKT8 cell percentage was found increased in both groups of patients with a decrease of OKT4 cell percentage in cerebral malaria patients only. Both groups thus exhibited a decreased OKT4 OKT8 ratio, which was slightly lower in cerebral malaria cases than in benign cases. NK activity was found elevated in uncomplicated cases of malaria, in contrast to patients suffering cerebral malaria, who exhibited a profound depression of NK activity.

Characterization of T Cells Cultured From Fine Needle Aspirates of Human Renal Allografts During Rejection

The fine needle aspiration (FNA) of cells infiltrating renal allografts has proven to be a useful and safe tool for monitoring renal graft outcome [1]. The recent availability of broad panels of murine monoclonal antibodies defining lymphocyte subsets has permitted the fine characterization of the profile of the FNA cell membrane. Thus, it has been shown that the onset of graft rejection is associated, in some cases, with the presence of large proportions of infiltrating cells that express the membrane antigen characteristic of suppressor cytotoxic lymphocytes (that is, OKT8 cells) [2, 31. It remains to be shown, however, whether these cells possess the functional capacities conesponding to this phenotype. Until now, this has been impossible due to the small number of cells recovered from FNA samples. The aim of this brief report is to describe a technique which allows the in vitro expansion of the lymphoid cell population contained in FNA samples in long-term cultures and thus the simultaneous analysis of their phenotypes and functions. Methods. Fine needle aspiration samples were obtained from six renal allograft recipients undergoing rejection. In all the patients the FNA was performed before the onset of antirejection treatment (that is, high dose corticotherapy). Freshly collected FNA samples were analyzed in parallel with blood samples drawn at the same moment from the same patients. In the present study only the aspirates containing high proportions of blastic mononuclear cells that were absent from peripheral blood were considered. Hemolysis of the red cells contaminating the cell samples was performed using the following .buffer: 8.29 g NH4C1, 37 mg disodium EDTA, and I g KHCO3 per liter, pH 7.3. Leukocytes were washed and recovered following a 10-mm centrifugation at 1500 rpm. The viable cells were counted by trypan blue dye exclusion, and cultured in round-bottomed microplates (Nunclon, Roskild, Sweden) (1 x iO to 2 x l0 cells per microwell in 0.2 ml). The number of cells recovered from FNA samples usually ranged from 5 x iO to 15 x i04 cells, depending on the patient. The conditioned medium (CM) used for culture was RPMI 1640 (GIBCO, Paisley, United Kingdom) supplemented with antibiotics, 50% crude supernatant containing significant levels of interleukin-2 (IL-2), 10% heatinactivated, fetal calf serum (FCS) (GIBCO, Paisley, United Kingdom), 5 x l0M mercaptoethanol and l%glutamine. The IL-2 containing supernatant was produced by stimulating normal human peripheral blood lymphocytes with phytohemagglutinin A (DIFCO, Detroit, Michigan, USA) for 48 hr as previously described [4]. The supernatants used to grow FNA cells contained approximately 0.7 to I U/ml of IL-2 activity (assessed by CTLL2 proliferation, see [5] for the definition of an IL-2 unit). After the initial 4 days of culture, the cells were fed every 48 to 72 hr by replacing half of the culture medium with fresh CM. Cell proliferation was regularly evaluated by means of cell counting with trypan blue dye, in parallel with 3H-thymidine incorporation. This latter method was particularly useful for the assessment of proliferation during the early days of culture. By day 4 of the culture, the FNA cells from all patients began to incorporate significant amounts of 3H-thymidine: patient 1: day 4,4177 cpm; day 5, 9042 cpm; day 6, 19335; patient 2: day 3, 548 cpm; day 4, 2353 cpm; day 5, 16506 cpm. Moreover, by days 7 to 11, depending on the patient, and approximately until day 20, the growth rate increased significantly with a doubling time of 6 hr (assessed by cell counting). Thereafter, the proliferation decreased with a doubling time close to 12 hr. All cultures were successfully maintained for 30 to 45 days. Phenotypic analysis of cultured cells was performed at regular intervals by means of an indirect immunofluorescence microtest, described by Cashman et al [6], using the OKT series of monoclonal antibodies. The antibodies used were OKT3 (IgG2a), OKT4 (IgG2b), and OKT8 (IgG2a) which react, respectively, with mature peripheral human T cells, helper/inducer T cells and suppressor/cytotoxic T cells [7]. In

T-cell subpopulations in patients with monoclonal gammopathies: essential monoclonal gammopathy, multiple myeloma, and Waldenstrom macroglobulinemia.

T-cell subsets defined by monoclonal antibodies (OKT3, OKT4, and OKT8) were analyzed in 117 patients with monoclonal gammopathies--69 multiple myeloma (MM) (30 untreated and 39 treated), 14 Waldenström's macroglobulinaemia (WM), and 34 essential monoclonal gammopathy (EMG) patients. The percentage and absolute numbers of total T-lymphocytes (E+, OKT3+ cells) were within the normal range in all groups except for the treated MM patients, in which a decrease in the absolute number could be observed. The percentages of OKT4+ cells were significantly lower in MM (35 +/- 1.7) than in EMG patients (43 +/- 2) and controls (50 +/- 2). In contrast, OKT8 cells correspondingly increased in MM (38 +/- 1.6) compared with EMG patients (29 +/- 1) and controls (27 +/- 1). The OKT4/OKT8 ratio was lower in MM than that in EMG patients and controls (p less than 0.01) and was shown to be one of the four most significant variables in a linear discriminant analysis used to distinguish between MM and EMG groups. The MM patients in clinical stage III as well as Bence-Jones myeloma patients showed a more pronounced OKT4/OKT8 imbalance. The treatment did not influence the percent distribution of T-cell subpopulations. The patients with WM exhibit an alteration in the distribution of the T-cell subsets similar to the MM patients with a T4/T8 ratio of 1.1 +/- 0.1. This imbalance was more pronounced in WM patients with monoclonal B-lymphocytes in peripheral blood (leukaemic phase of WM). The functional significance of the altered T-cell subsets in MM and WM patients remains to be established, though it is probable that such an imbalance plays an important role in regulating these B-cell proliferations.

Bronchoalveolar lavage cells and proteins in patients with the acquired immunodeficiency syndrome. An immunologic analysis.

Several components of cellular and humoral immunity were examined in bronchoalveolar lavage fluid and blood of 15 patients with the acquired immunodeficiency syndrome, and the results were compared to data from 25 healthy controls (including 5 asymptomatic homosexual men). Compared with that of controls, bronchoalveolar lavage fluid from patients tended to have more lymphocytes and significantly more neutrophils; a lower OKT4/OKT8 ratio, due to an increase in total OKT8 cells; and normal total OKT4 cell counts, despite a significant decrease in numbers of OKT4 cells in peripheral blood. Patients also had significantly more IgG-releasing cells and higher IgG levels than controls in lavage fluid. These data show that, in the lung lining fluid of patients with the acquired immunodeficiency syndrome, significant alterations in cellular and humoral immunity exist that differ in several important respects from immunity in controls and from corresponding changes in patients' peripheral blood.

COMPARISON OF SURFACE DIFFERENTIATION ANTIGENS BETWEEN SEPARATED HGPRT-POSITIVE AND NEGATIVE T-CELL POPULATIONS FROM TWO LESCH-NYHAN HETEROZYGOTES: 233

Peripheral T-lymphocyte populations from two Lesch-Nyhan heterozygotes were found to consist of 95% hypoxanthineguanine phosphoribosyltransferase (HGPRT)-positive and 5% negative cells by the culture of the T-cells with 6-thioguanine. The T-cells frome each heterozygote were cultured in the conditions in which either enzyme-positive (azaserine-hypoxanthine medium) or negative (6-thioguanine medium) cells selectively proliferate. After two weeks, each separated T-cell subpopulation was considered to be pure with regard to the enzyme expression. Bewteen the enzyme-positive and negative subpopulations, percentages of OKT3, OKT4 or OKT8-positive T-cells were compared. The fact that only small percentage of peripheral T-cells are HGPRT-negative in the heterozygotes suggests that in vivo differentiation of HGPRT-negative cells is disturbed in the heterozygotes. Our data have shown that both enzyme-positive and negative T-cell subpopulations from each heterozygote possessed equivalent percentages of OKT4, OKT8 and OKT3-positive cells. These results suggest that, in the heterozygotes, growth or differentiation of HGPRT-nnegative lympoid cells of T-cell linkage was disturbed before OKT4 and OKT8-bearing thymocyte subpopulations separate from each other.