Percentage Of OKT8 Research Articles

Studies on subsets, NK and ADCC activity of lymphocytes in synovial fluids of patients with rheumatoid arthritis

Subsets of lymphocytes in synovial fluids of patients with rheumatoid arthritis (RASF-Ly) along with lymphocytes in peripheral bloods of normal subjects (Healthy-Ly) were studied.Percent of OKT-3+cells (P<0.01), OKT-4+cells (P<0.01), OKT-8+cells (P<0.05) and OK-Ial+cells (P<0.01) of RASF-Ly were significantly higher than those of Healthy-Ly. Therefore, we examined the correlation these percentage of subsets. Positive correlation were observed in RASF-Ly between percentage of OKT-8+cells (P<0.01) or Leu-7+cells (p<0.01) and that of OK-Ial+cells and also between percentage of OKT-8+cells and that of Leu-7+cells of RAST-Ly (p<0.01) . Since OK-Ial represents a marker of activated T cells, both OKT-8+cells and Leu-7+cells in RASF-Ly seemed to be activated and Leu-7+cells seemed also to be immature. NK activity of RASF-Ly was significantly lower than that of Healthy-Ly (p<0.01), whereas ADCC activity of RASF-Ly was similar to that of Healthy-Ly.

Influence of operative trauma on circulating blood mononuclear cells: analysis using monoclonal antibodies.

The influence of operative trauma on the quantity and proportion of cooperating mononuclear cells in peripheral blood of patients with stage 0 uterine cancer who underwent hysterectomy was studied using monoclonal antibodies. The percentage of OKM1+ cells (monocytes) had increased on day 1 by 12.4%, on day 3 by 24.8%, and had returned to normal by day 7. The percentage of large OKIa1+ cells (expressing Ia antigen) on day 1 was increased by 186.9%, while on day 3 it had returned to a normal level. There was a decrease on day 1 of the number and percentage of OKT3+ cells (all T lymphocytes) by 17.4%, OKT4+ cells (helper/inducer cells) by 27.9%, OKT8+ cells (suppressor/cytotoxic cells) by 24.6%, and small OKIa1+ cells by 49.4%. The number and percentage of T lymphocytes remained below the preoperative level throughout the observation period. However, the number of helper cells had increased significantly on day 5, while suppressor cells remained below the preoperative level during the entire observation period. The rising ratio of OKT4+/OKT8+ cells reflected the kinetics of recovery of the helper over the suppressor cell population.

Mycosis fungoides

Peripheral blood lymphocytes from 16 mycosis fungoides (MF) patients were studied using OKT series monoclonal antihuman T cell antibodies. The percentage of OKT3+ cells was in normal range for all MF patients compared with controls; the percentage of OKT4+ cells was significantly increased (p &lt; 0.002) in MF patients versus controls; the percentage of OKT8+ and OKT11+ cells in the MF group did not differ from controls. The OKT4+ cell expansion was apparently not dependent from the clinical stage of disease. These findings suggest that in MF patients there is a circulating OKT4+ cell expansion and, indirectly, that MF could be regarded as a helper T cell neoplasm.

Détermination des sous-populations de lymphocytes circulants dans la dermatite atopique à l’aide d’anticorps monoclonaux

Monoclonal antibodies were used to determine the level of circulating helper and suppressor T cells in 34 infants and adults with severe atopic dermatitis and in normal controls. The percentage of OKT3 (total T lymphocytes) was reduced significantly in all the atopic infants. The percentage of OKT8 (suppressor-cytotoxic T lymphocytes) was reduced significantly in all patients with active lesions. The percentage of T gamma lymphocytes was reduced in all the atopic patients with or without active lesions.

Correlation of plasma hormone levels and peripheral circulating lymphocyte subpopulations during human pregnancy

Using monoclonal antibodies (OKT3, OKT4, OKT8) peripheral blood lymphocyte subsets were determined in 40 normal primiparous pregnant women and compared with those of 31 nonpregnant controls. In pregnant women plasma concentrations of estradiol, progesterone, human placental lactogen (HPL), beta subunit of human chorionic gonadotropin (βHCG), and α-fetoprotein were measured by means of radioimmunoassay. We studied if correlations between peripheral lymphocyte subsets and plasma hormone levels might exist. We observed in pregnant women from 10 to 40 wk of amenorrhea a decrease in the percentage of OKT3 and OKT8 cells, and during the course of pregnancy an increase in the percentage of OKT4 cells. This increase inversely correlated with plasma βHCG levels and directly correlated with plasma HPL levels.

Thymic lymphocyte subpopulations in myasthenia gravis.

We compared the percentages of thymic mononuclear cells (TMC) that bind monoclonal antibodies to T-cell subpopulations in patients with myasthenia gravis (MG) and non-MG patients undergoing cardiac surgery (CS). There were no significant differences in percentages of OKT3+, OKT4+, OKT6+, or OKT8+ cells or the OKT4:OKT8 ratio. There was an increase in the percentage of Ia+ (immune response gene-associated antigen) TMC in MG compared with CS but no significant differences in B cells or phagocytic cells. The Ia+ cells could be abnormal B cells, activated T cells, or thymic dendritic cells.

Mechanisms of T-cell unresponsivenessin leprosy

Summary We analysed the mechanisms of T-cell unresponsiveness to Mycobacteriumleprae antigens and to unrelated antigens or T-cell mitogens in human leprosy and in an experimental model of murine infection by M. lepraemurium (MLM). In human leprosy, monoclonal antibodies OKT3, OKT4 and OKT8 were used to enumerate T-cell subpopulations within peripheral blood. Increased percentages of OKT8+ cytotoxic/suppressor cells were observed in untreated, non-reactional lepromatous patients. Conversely, lepromatous patients suffering from erythema nodosum leprosum, an Arthus-like phenomenon, exhibited a transient drop in the percentage of OKT8+ cells with a correlative increase in the proliferative response to T-cell mitogens. We studied the proliferative response to M. leprae of OKT4+ and OKT8+ cells isolated by a negative selection procedure using antibody-induced cytotoxicity plus complement. None of these subpopulations proliferated when incubated with M. leprae. In some patients, control treatment of mononuclear cells with complement alone induced the reappearance of a strong proliferative response to M. leprae, suggesting the existence of an active suppressor mechanism through soluble factors of an unknown nature. In MLM-induced murine leprosy, a progressive decrease was observed in the proliferative response to concanavalin A (ConA), and an early decrease in interleukin 2 activity in supernatants from ConA-stimulated spleen cells. Splenic T cells from MLM-infected mice transferred into naive recipients accelerated the local MLM growth in these recipients, suggesting that suppressor T cells may play a pathogenic role in the progression of MLM infection.

T-cell subsets in human lymphocytes maintained in IL-2 medium after PHA or mixed lymphocyte reaction activation

The culture of human T lymphocytes in interleukin-2 (IL-2) containing growth factor medium results in a significant shift in the T-lymphocytes subsets isolated from such cultures at weekly intervals. If normal peripheral blood mononuclear cells are stimulated with phytohemagglutinin (PHA) or in a mixed lymphocyte reaction (MLR), the resulting T lymphoblasts can be propagated in growth factor medium. Staining of the cultured cells with monoclonal antibodies was evaluated by indirect immunofluorescence on a laser-activated flow cytometer (Ortho Spectrum III). The antibodies used were: OKT3 (mature T lymphocytes), OKT4 (helper/inducer T lymphocytes), OKT8 (cytotoxic/suppressor T lymphocytes, OKT10 (immature and “activated” lymphocytes). OKT11a (cells which rosette with sheep erythrocytes), and OKIa-I (HLA-DR constant region). Both PHA and MLR activation resulted in initial preservation of the OKT4+ subset predominance over OKT8+ T lymphocytes noted on normal circulating blood lymphocytes. However, during culture in T-cell growth factor medium, there was a progressive increase in the percentage of OKT8+ cells, and a concomitant decrease in OKT4+ lymphoblasts. The increase in OKT8+ cells in the MLR-stimulated cultures was paralleled by an increase in specific cell-mediated cytotoxicity against the stimulating lymphocyte population. In addition to the shift in T-lymphocyte subset, there was virtual 100% staining with OKT3 and OKT11a, indicating the T-cell nature of the proliferating cells. OKT10 which was present on a small subset of fresh blood lymphocytes appeared rapidly in stimulated cultures, and was retained on virtually all lymphoblasts of either OKT4+ or OKT8+ subset, OKIa-1 cells increased slowly in PHA-stimulated cultures. HLA-DR+ T cells were detected earlier in MLR cultures. The activation of T lymphocytes results in a significant increase in the number of molecules of OKT11a bound per cell, in concert with the increased avidity of T lymphoblasts for sheep erythrocytes. The significant change in the phenotype and function of lymphoblasts isolated from long-term cultures demonstrates the importance of monitoring cultures, and the potential hazards in equating a cultured cell population with a freshly isolated one.

Functional analysis of monkey lymphocyte subsets defined by OKT4 and OKT8 monoclonal antibodies

The percentages of rhesus monkey blood lymphocytes (PBL) reactive with OKT4 and OKT8 antibodies and the OKT4 OKT8 ratio showed significant correlations with the log of the immunoglobulin plaque-forming cell (PFC) response after stimulation with pokeweed mitogen (PWM). These correlations suggested that monkey OKT4 + cells function as “helper” cells and OKT8 + cells function as “suppressor” cells for the PFC response. This was confirmed by separation and study of enriched T- and B-cell subpopulations. OKT8-depleted (OKT4 +) and OKT4-depleted (OKT8 +) cells were obtained by treatment of purified T cells with antibody and complement. OKT4 + cells augmented the PWM-induced B-cell differentiation into PFC but OKT8 + cells did not. OKT8 + cells suppressed the PFC response by mixtures of B cells and OKT4 + cells. OKT8 antibodies also detected a suppressive cell subset in African green monkeys since the percentage of OKT8 + cells showed a negative correlation with the log PFC response. OKT4 antibodies failed to bind to African green monkey PBL.

T-cell dysfunctions in IgA nephropathy: Specific abnormalities in the regulation of IgA synthesis

This work was undertaken to determine the cellular abnormalities that could explain the high levels of serum IgA frequently found in patients with IgA nephropathy. Seventeen control subjects and twenty-seven patients who had received no therapy were studied. After in vitro pokeweed mitogen (PWM) stimulation, significantly higher amounts of IgA were produced by peripheral blood mononuclear cells (PBM) of patients when compared with those of the control group (560 ± 97 vs 231 ± 57 ng/ml, P < 0.0025). No differences were observed in the synthesis of IgG and IgM. Twenty out of twenty-seven patients presented an increase in the percentages of OKT4 + cells (mean + 2 SD), in relation to the control group, with normal or elevated percentages of OKT8 + cells. The OKT4 + OKT8 + cell ratio was elevated in 12 out of 27 patients. All patients presented some abnormality in the generation of IgA-specific suppressor cells at variable doses of concanavalin A (Con A) on in vitro PWM-stimulated culture of PBM. In both assays low doses of Con A (2.5 μg/ml) induced a certain suppression of IgA synthesis in patients that was not observed in the majority of the control group. At these doses some patients also showed an enhancement in the synthesis of IgG and IgM. On the contrary, higher doses of Con A (50 μg/ml) produced significantly less IgA suppression than the controls. Normal IgA-suppression values were found at 10 μg/ml of Con A. T cells obtained from patients were significantly more efficient than T cells from controls in providing IgA-helper activity for normal allogeneic enriched B cells ( P < 0.025) in PWM-stimulated cocultures. These results show that patients with IgA nephropathy present, after mitogen stimulation in vitro, a specifically increased production of IgA as well as an augmentation in the activity of IgA-helper T cell and a deregulation on IgA-suppressor T-cell function. According to these data, it is suggested that the alteration observed in helper T cells might precede that of suppressor T cells. These immunoregulatory abnormalities might contribute to the pathogenesis of the disease.

Reversible Alterations in Immunoregulatory T Cells in Smoking: Analysis By Monoclonal Antibodies and Flow Cytometry

We characterized T-lymphocyte subsets in peripheral blood of smokers (N = 60) and nonsmokers (N = 35). Total T-lymphocytes and T cell subsets were similar to nonsmokers in light and moderate smokers. In heavy smokers, total OKT3+ cells were increased, the percentage of OKT4+ cells was decreased, and percentage and total number of OKT8+ cells were increased. The ratio of OKT4+ to OKT8+ lymphocytes was decreased in heavy smokers. The percentage of OKT8+ cells and the OKT4+/OKT8+ ratio returned to normal in heavy smokers six weeks after they stopped smoking. These findings suggest that cigarette smoking causes reversible alterations in immunoregulatory T cells.

T-lymphocyte subsets in peripheral blood and lung lavage in idiopathic pulmonary fibrosis and sarcoidosis: Analysis by monoclonal antibodies and flow cytometry

Thymus-dependent (T) lymphocytes may contribute to the pathogenesis of human interstitial lung disease. In order to determine whether alterations of immunoregulatory T cells occur in patients with idiopathic pulmonary fibrosis (IPF) and sarcoidosis, we characterized T lymphocytes in peripheral blood ( n = 8 and 11, respectively) and lung lavage ( n = 4 and 6, respectively) in untreated patients with these diseases. In IPF, we found a decreased percentage, but normal total count of circulating OKT3 + (mature) and OKT4 + (inducer/helper) cells compared to normal controls. We observed a normal percentage and total count of circulating OKT8 + (cytotoxic/suppressor) cells. The ratio of OKT4 + to OKT8 + ( 4 8 ) lymphocytes, reflecting the balance of immunoregulatory cells, was normal in peripheral blood. Comparing peripheral blood to lung lavage, we noted a lower proportion of OKT4 + cells and a higher proportion of OKT8 + cells in lung lavage. The 4 8 ratio in lung lavage tended to be low compared to blood. In contrast, we found in sarcoidosis patients a decrease in both percentage and total circulating OKT3 +, OKT4 +, and OKT8 + cells as compared to normal. In lung lavage, there was an increase in OKT3 + cells, due to an increase in the OKT4 + subset. The percentage of OKT8 + cells in lung lavage was low. Compared to blood the 4 8 ratio was high in lung lavage. Thus, a number of alterations in circulating and lavage T cells were found both in patients with IPF and sarcoidosis. These results suggest that immunoregulatory abnormalities contribute to pathogenesis of these disorders.

Reduction of the Fraction of Circulating Helper-Induced T Cells Identified by Monoclonal Antibodies in Psoriatic Patients Treated with Long-term Psoralen/Ultraviolet-A Radiation (PUVA)

Ultraviolet radiation has been found to alter the distribution and function of human lymphocytes. To determine whether photochemotherapy (PUVA) alters circulating levels of T cell subset marker-bearing lymphocytes, cells from 9 patients with psoriasis undergoing PUVA therapy for several years (mean 4.6 +/- 1.4 yr), 17 patients with active untreated psoriasis, and 20 healthy volunteers were reacted with monoclonal antibodies to T cell surface markers, including OKT3 (all peripheral blood T cells), OKT4 (helper/inducer T cells), OKT6 (common thymocytes), and OKT8 (suppressor/cytotoxic T cells), and analyzed by flow cytometry. There were no differences in the distribution of T cell subsets between healthy volunteers and patients with active psoriasis. In contrast, the percentages of lymphocytes reacting with OKT3 and OKT4 were lower (by 16% and 12% percent respectively, p less than 0.0025) in the PUVA-treated patients compared to healthy volunteers or patients with active psoriasis that had not received PUVA therapy. There was no difference in the percentage of OKT8 and OKT6 bearing cells. Squamous cell carcinoma of the skin subsequently developed in 2 of 3 PUVA-treated patients with the lowest percentages of T4-bearing cells. These findings indicate that long-term PUVA therapy is associated with a reduction in circulating helper/inducer T cells. This reduction may have a role in the altered immune function reported in PUVA-treated patients.

The Expression of Surface Antigen Leu 1 by Ocular Adnexal Lymphoid Neoplasms

AbstractLeu 1 is a surface membrane antigen expressed by benign systemic T cells, most cases of B-cell-derived chronic lymphocytic leukemia, and approximately one half of the systemic B-cell lymphomas. Benign systemic B cells are Leu 1-. Our studies showed that the percentage of Leu 1+ cells was essentially the same as the percentage of OKT3+E+ (T) cells in six ocular adnexal pseudolymphomas, strongly suggesting that the benign B cells in these lesions were similarly Leu 1-. These studies also showed that malignant B cells in six of 15 (40%) ocular B-cell lymphomas were Leu 1+. Thus, the expression of Leu 1 by some malignant ocular adnexal B cells but not by benign ocular adnexal B cells appears to be analogous to the situation for lymph nodal and other systemic B cells. Although the explanation for the existence of this shared determinant is unknown, Leu 1 expression may possess clinical and diagnostic significance as a marker of neoplastic B cells

T cells in monoclonal gammopathies.

Subpopulations of human T lymphocytes were analysed by monoclonal antibodies (OKT3, OKT4, OKT8) in healthy controls and in patients with multiple myeloma (MM), Waldenström's macroglobulinemia (WM) and benign monoclonal gammopathy (BMG). Lymphocytes forming rosettes with SRBC correlated well with T cells staining in indirect immunofluorescence by OKT3 monoclonal antibodies. The relative and absolute numbers of OKT4+ T cells were significantly lower in patients than in controls. Though, the percentage of OKT8+ T cells was increased in patients, the total OKT8+ cell counts were normal in all patient groups. The ratio between OKT4+ and OKT8+ lymphocytes was low in the groups of treated MM and of WM patients compared to controls (P less than 0.001). Moreover, the ratio was lower than the normal range in 27% of BMG and 38% of untreated MM patients. The imbalance between OKT4+ and OKT8+ T cells in untreated MM was more pronounced in clinical stage III patients than in stage I and II patients. The most pronounced changes were noted in treated MM patients. The significance of the altered T cell subsets in monoclonal gammopathies with regard to polyclonal and tumor B cell regulation remains to be established.

Monoclonal antibody analysis of blood T-cell subsets in myasthenia gravis.

Analysis of peripheral blood T -cell subsets and B-cells in patients with myasthenia gravis was performed using monoclonal antibodies and antibody against surface immunoglobulins (SIg) in an immunofluorescent technique. We found a modest but significant decrease in percentages of OKT3- and OKT8-positive cells (thought to represent total T-cells and T-suppressor/cytotoxic cells, respectively) in myasthenics as a group. The percentage of OKT3-positive cells was significantly decrease in patients with late-onset disease (greater than 35 years old), while the percentage of OKT8-positive cells was significantly reduced in those with early-onset myasthenia (greater than 35 years old). Both thymectomized and nonthymectomized patients exhibited a decreased percentage of OKT3-positive cells. No significant difference was found between the percentages of SIg-positive cells in myasthenics and controls. Our results suggest that only modest imbalances of circulating immunoregulatory lymphocytes occur in myasthenia gravis; however, it is conceivable that the small differences observed in this study may reflect pathogenetically important reductions in a functionally distinct lymphocyte subpopulation.

The measurement of leukocyte subsets in the peripheral blood of cancer patients with solid tumors using monoclonal antibody reagents.

Monocyte and lymphocyte subsets were quantitated in the peripheral blood of normal subjects and patients with solid tumors using the monoclonal antibody reagents OKM1, BRL63D3, OKT3, OKT4, and OKT8. Percentages and numbers of cells reacting with monoclonal reagents were analyzed by indirect immunofluorescence. Correlations between leukocyte subset values and stage of disease or immunocompetence were sought. No differences from normal were seen in the percentage of OKT3, OKT4, and OKT8 cells or in the ratios of OKT4/OKT8 cells in peripheral blood mononuclear cells (PBMC) from all cancer patients, patients with localized disease, or patients with advanced disease. A significant decrease in absolute numbers of lymphocytes, OKT3 cells, OKT4 cells, and OKT8 cells was seen in the peripheral blood of patients with advanced disease reflecting the absolute lymphopenia of these patients. A significant increase was seen in the percentages of PBMC reacting with OKM1 and BRL63D3 from patients with advanced disease compared with normal donors or localized disease patients. A positive correlation was found between PHA responsiveness and absolute numbers of OKT3 and OKT4 cells. A negative correlation was found between PHA responsiveness and percentages of OKM1 cells. These data indicate that malignant disease does not alter T cell subset percentages in patient peripheral blood but may decrease their absolute numbers in association with absolute lymphopenia. On the other hand, percentages of OKM1 and BRL63D3 cells are increased in patients with advanced solid tumors in association with impaired PHA responsiveness.

Interstitial mononuclear cell populations in renal graft rejection. Identification by monoclonal antibodies in tissue sections.

The interstitial mononuclear cell populations of 22 renal grafts with interstitial rejection (IR), 6 grafts with interstitial nephritis without rejection (IN), and 5 kidneys without infiltration (3 donor kidneys, 2 grafts) were identified and quantitated by monoclonal antibodies recognizing T cells (TA-1, OKT3), helper inducer cells (OKT4), cytotoxic/suppressor cells (OKT8), B cells (BA-1), and monocytes and null cells (OKM1). Double-layer fluorochrome enhancement using F(ab')(2) reagents and nuclear counter staining with ethidium bromide enabled quantitation of the number of positive mononuclear cells, interstitial cells, and total cells on each of 30-55 microscopic fields per tissue section. T cells were the most abundant infiltrating cell in tissues with IR (35 +/- 9.8 percent), significantly higher than that seen in IN (21 +/- 16 percent) or in kidneys without infiltration (5.0 +/- 3.9 percent). The percentage of T cells identified by TA-1 or OKT3 was approximately equivalent to the summation of OKT4 plus OKT8. Although no differences were observed in the percentage of OKT4 cells, the percentage of OKT8 was significantly higher in IR (26 +/- 7.7 percent, P {less than} 10(-4)) than in IN (9.3 +/- 6.2 percent) or in kidneys with normal interstitium (3.0 +/- 2.4 percent). The ratio of OKT8/OKT4-positive T cells in 22 graft tissues with IR (3.2 +/- 1.4) was greater (P {less than} 0.0007) than 6 graft tissues with IN without rejection (0.82 +/- 0.39) and the 5 kidney tissues without interstitial infiltration (0.75 +/- 0.25). There was no significant difference between the groups in the relatively low percentage of interstitial cells identified as B cells reacting with BA-1 or containing S(IgD,M). The percentage of interstitial cells recognized by OKM1 was similar in rejection and interstitial nephritis, with both being greater than controls (P {less than} 0.02). The relative numbers of blood mononuclear cells identified by the monoclonal antibodies was generally not predictive of the proportions present in kidney tissue, although OKT4-positive blood cells were less numerous and OKMI+ blood cells were more numerous than in controls (P {less than} 0.002). Quantitative analysis of identifiable interstitial cells in graft rejection reveals that most infiltrating cells are T cells, the greater proportion of which are recognized by OKT8. OKT8-positive cells may play an important role in mediating renal graft rejection.

Numeration of T cell subsets in sarcoidosis using monoclonal antibodies: decreased levels of peripheral blood T cells and cells with suppressor T cell phenotype.

Peripheral blood lymphocytes frm 16 patients with sarcoidosis (9 patients with skin sarcoids, 7 patients with erythema nodosum and bilateral hilar adenopathy) and cutaneous anergy and from 23 age-matched healthy controls were characterized by reactivity with monoclonal antibodies OKT3, OKT4, OKT8 directed to surface antigens of T lymphocytes, helper-inducer and suppressor-cytotoxic T cell subsets, respectively. In contrast to healthy controls, patients with sarcoidosis had reduced percentages of OKT3+, OKT4+ and OKT8+ cells and a major decrease in the OKT8+ (suppressor) subset. However, these changes were significant only in the group of patients with acute sarcoidosis (erythema nodosum). This abnormal T cell distribution correlates with the alterations in cell-mediated immunity previously observed and suggests the presence of a defective circulating suppressor T cell activity in acute sarcoidosis.