OKT8-positive Cells Research Articles

Immunocytological study of phlyctenular eye disease.

Scrapings from phlyctens and conjunctiva of 12 patients with phlyctenular keratoconjunctivitis were studied using OKT4-Leu3a, OKT8, B1, BA1, S-100 and HLA-DR monoclonal antibodies. T-lymphocytes were present in both conjunctival and phlyctenular scrapings. OKT4-Leu3a positive cells outnumbered the OKT8 positive cells in both conjunctival and phlyctenular scrapings. B1 and BA1 positive cells were absent from the conjunctival scrapings, but were present in the phlyctenular scrapings. S-100 positive cells were present in both conjunctival and phlyctenular scrapings. However, they were very few in the conjunctival scrapings. Most of the cells in both conjunctival and phlyctenular scrapings were HLA-DR positive. These findings support the hypothesis that cell mediated immunity is responsible for the pathogenesis of phlyctenular eye disease.

The behaviour of OKT3-, OKT4- and OKT8-positive cells during phases of elevated spontaneous chemiluminescence activity (CL-A) in multiple sclerosis patients. A serial study.

The chemiluminescence activity (CL-A; synonym = burst activity, BA) and the percentage of OKT3-, OKT4- and OKT8-positive peripheral blood cells were serially examined in four control persons and in eight patients with multiple sclerosis. When the OKT values obtained in phases of increased CL-A (clinical remission) were compared with those of the control group, the percentage of OKT3-positive cells was reduced (P = 0.014), and that of OKT4-positive cells increased (P = 0.014); there were no significant changes in the percentage of OKT8-positive cells (P = 0.171). After the CL-A had returned to normal values, the OKT4-positive cells remained elevated (P = 0.029), whereas the OKT3- (P = 0.342) and OKT8-positive cells (P = 0.443) showed no significant changes. When in the same patients, phases of increased CL-A were compared with phases in which values were not elevated, a reduced percentage of OKT3-positive cells was found in phases with increased CL-A (P = 0.031); however, the OKT4-positive and OKT8-positive cells did not differ significantly (P = 0.156 and 0.281).

ＩＢＭ２９９７を用いた血小板およびか粒球採取時の供血者ならびに採取バッグ中のリンパ球サブセット

In the platelet apheresis and granurocyte apheresis used by the cell-separater: IBM 2997, the changes of absolute number of lymphocytes and lymphocyte subsets were investigated in comparison with pre- and post-apheresis in the peripheral blood of donors. Also, lymphocyte subsets into the bags collected from donors were examined.In the platelet apheresis, absolute number of lymphocytes and precentages of lymphocyte subsets were not changed in the peripheral blood of donors.However, in the granulocyte apheresis, after injection of hydrocortizone (200mg), percentages of lymphocyte, OKT3 and OKT4 positive cells in the peripheral blood of donors were decreased significantly compared to pre-apheresis.On the other hand, OKT4/8 ratio into bags collected by granulocyte was more decreased significantly compared to platelet apheresis bags.From these results, we strongly feel that the prevention of lymphocyte contamination and calculation of OKT4/8 ratio into the collected bags will be important to avoid the allergic reaction and GVHD.

Immunohistochemical study of Langerhans cells in skin tumors.

The behavior of Langerhans cells in skin tumors was investigated by immunohistochemical techniques using OKT6. OKT6-positive cells were numerous in squamous cell carcinomas, seborrheic keratoses and keratoacanthomas. They were rare in basal cell carcinomas, Bowen's disease, eccrine poromas, extramammary Paget's disease and warts. In solar keratosis, the number of OKT6-positive cells was almost equal or slightly larger compared to the normal epidermis. These results indicate that the density of Langerhans cells may not correlate with the degree of malignancy but, to a certain extent, with the nature of the membrane of tumor cells occurring e.g. during keratinization.

Polymyositis Associated With AIDS Retrovirus

Two homosexual men were initially seen with polymyositis as the only manifestation of the acquired immunodeficiency syndrome (AIDS) retrovirus infection. They developed AIDS-related complex a few weeks later and typical AIDS two to six months after onset of muscle weakness. By use of anti-human T-cell lymphotropic virus type III antiserum and monoclonal antibodies to lymphocyte subsets in an immunofluorescence technique, viral antigens were found in the OKT4-positive lymphoid cells surrounding muscle fibers and invading the endomysia septa. We concluded that an initial infection with the AIDS retrovirus can be associated with polymyositis, which may be the first clinical manifestation of an impending AIDS-related complex or AIDS.

Polymyositis associated with AIDS retrovirus

Two homosexual men were initially seen with polymyositis as the only manifestation of the acquired immunodeficiency syndrome (AIDS) retrovirus infection. They developed AIDS-related complex a few weeks later and typical AIDS two to six months after onset of muscle weakness. By use of anti-human T-cell lymphotropic virus type III antiserum and monoclonal antibodies to lymphocyte subsets in an immunofluorescence technique, viral antigens were found in the OKT4-positive lymphoid cells surrounding muscle fibers and invading the endomysia septa. We concluded that an initial infection with the AIDS retrovirus can be associated with polymyositis, which may be the first clinical manifestation of an impending AIDS-related complex or AIDS. (<i>JAMA</i>1986;256:2381-2383)

Activated T cells in normal pregnant women and neonates

The phenotyping of T-cell subsets and T cells at different stages of activation was performed using a panel of monoclonal antibodies in samples from normal pregnant women at different stages of gestation and in the cord blood of neonates. The data obtained from pregnant women showed a slight decrease in the total number of T cells at the beginning of pregnancy, whereas there was a clear increase in 4F2-positive lymphocytes after a few months of gestation. No significant increase in Class II-positive lymphocytes was observed in normal pregnant women in comparison with adult healthy women. The data from neonates revealed a clear decrease of OKT3- and OKT4-positive cells and an increase of 4F2-positive cells in comparison with control subjects. These data indicate that alerted, but not fully activated, lymphocytes are present in the circulation of both the mother, after the first months of pregnancy, and the neonate. This finding reinforces the concept that during pregnancy there is an activation of certain immune components rather than a general depression of the immune system.

Human genital papilloma infections: An evaluation of immunologic competence in the genital neoplasia-papilloma syndrome

Immunologic evaluations of women with genital neoplasia-papilloma syndrome demonstrated the presence of subclinical immunodeficiency when compared with results in 20 control women. All patients with genital neoplasia-papilloma syndrome were previously found to have human papillomavirus deoxyribonucleic acid in genital neoplasias or papillomas occurring either synchronously (in at least two genital organs at the same time) or metachronously (at different times during a period of months to years). Immunologic tests included blastogenic responses of lymphocytes to mitogens (phytohemagglutinin, concanavalin A, pokeweed mitogen, and tetanus antigen) and lymphocyte phenotyping with the use of monoclonal antibodies (OKT3, OKT4, OKT8, and OKT11). As compared with those of control subjects, the responses of the lymphocytes of patients with genital neoplasia-papilloma syndrome to mitogens were significantly decreased. The group with genital neoplasia-papilloma syndrome had a significantly higher percentage of suppressor-cytotoxic T cells (OKT8-positive cells) when compared with that of control subjects (mean 33% versus 18%) and a lower proportion of helper T cells (OKT4-positive cells) when compared with that of control subjects (35% versus 50%). The mean helper-to-suppressor/cytotoxic T-cell ratio (mean OKT4/OKT8 ratio) in the human papillomavirus-infected women was 1.72 +/- 0.29 (SE) as compared with 3.21 +/- 0.33 (SE) in the control group, demonstrating a significant reduction of the ratio in the patients with genital neoplasia-papilloma syndrome. These findings suggest that patients with genital neoplasia-papilloma syndrome have a reduced suppressor/cytotoxic T-cell ratio (mean OKT4/OKT8 ratio; that in the human papillomavirus-infected women was 1.72 +/- 0.29 (SE) as compared with 3.21 +/- 0.33 (SE) in the control group, demonstrating a significant reduction of the ratio in patients with genital neoplasia-papilloma syndrome. These findings suggest that patients with genital neoplasia-papilloma syndrome have reduced immunocompetence of unknown etiology.

Monoclonal antibody identification of infiltrating mononuclear leukocytes in lupus nephritis

Populations of mononuclear inflammatory cells infiltrating the renal interstitium in LN were studied by means of an avidin-biotin immunoperoxidase technique applied to cryostat sections of 26 renal biopsies (3 WHO class IIb; 4 class III; 8 class IV; 4 class V; 4 class III and V; and 3 class IV and V). The majority of interstitial leukocytes were T cells (mean 65.7 +/- 14.1). The number of cells reactive with OKT8 (47.3 +/- 11.0) exceeded the number of OKT4 positive cells (32.5 +/- 11.3) in 22 of 26 biopsies. Cells reactive with antimonocyte antibodies OKM1 and OKM5 (6.7 +/- 5.9 and 7.9 +/- 5.9, respectively) and B lymphocytes (OKB2 3.9 +/- 3.5) were a minor component of the interstitial infiltrates. Monocytes were the predominant cell type among stained cells in glomerular tufts and crescents. Tissue T4/T8 ratios varied widely (range 0.31 to 1.81), and were less than 1 in 22 of 26 patients. There was no correlation between tissue T4/T8 ratios and simultaneous peripheral blood T4/T8 ratios. Using stepwise multivariate linear regression, tissue T4/T8 ratio was found to correlate highly with renal histologic activity (P less than 0.001) but was not independently predictive of any other histopathologic or clinical variable studied. Mean tissue T4/T8 ratio in LN was significantly lower than that of other glomerular and interstitial diseases studied (P less than 0.001), a finding which may reflect differences in the pathogenesis of renal injury. These findings suggest that local cellular immune mechanisms may be important in the modulation of disease activity in LN.

Distribution of circulating mononuclear cells in short-term PUVA-treated psoriatic patients and healthy subjects

Peripheral blood mononuclear cells (PBMC), as defined by monoclonal antibodies (OKT3, OKT4, OKT8, OK 1, Leu 7, Leu 11b) were determined in 10 psoriatic patients and in 10 healthy subjects before and after administration of short-term PUVA therapy. A comparison of the mean baseline percentages of the two groups showed a statistically significant increase in Leu 7+ cells (p less than 0.001) as well as a slight increase in OKM1 and OKT8 positive cells in the psoriatic subjects. After 21 exposures, these subsets showed a reduction towards control values, while a significant increase in OKT3 and OKT4 positive cells (p less than 0.01) could be observed only in the control group. These results indicate that short-term PUVA therapy is associated with changes in PBMC subpopulations. This modification, however, does not necessarily imply a disturbance of immune system function, including natural killer activity.

Comparative phenotypic and ultrastructural characteristics of OKT6-positive cells in normal peripheral blood (adult and infant), in cord blood and in epidermis

Simple and rapid procedures have been developed to isolate T6-positive cells in epidermal cell suspensions, in peripheral blood of adult, and in cord blood. The isolated T6-positive cells were tested by double immunogold labelling at ultrastructural level to analize their phenotype (T6/HLA-DR antigens) and their morphological features in order to understand their origin, their fate, and their relationship to a) the early precursors of dentritic cells, b) immature neonatal lymphoid subsets, and c) immature macrophagic leukocytes.

Study of T cell subsets in patients with cutaneous leishmaniasis.

The proportion of T lymphocyte populations was assessed using monoclonal antibodies (OKT3, OKT4, OKT8) and spontaneous E-rosette tests with SRBC (for total and active TE cells) in 36 patients with cutaneous leishmaniasis subdivided according to the clinical variety. The T cell profile was normal in patients with simple cutaneous leishmaniasis of early ulcerative type whereas active TE cells and OKT4 positive cells were reduced in some cases of late ulcerative type. A significant reduction of active TE cells and of OKT4 positive cells (helper/inducer phenotype) and increase of OKT8 positive cells (suppressor/cytotoxic phenotype) was observed in peripheral blood of all patients with persistent active lesions and leishmaniasis recidivans. In patients with highly ulcerated persistent lesions a low proportion of active TE cells was also demonstrated in cellular extracts from dermal tissue. The results support the data obtained in experimental leishmaniasis that lack of helper/inducer cells and generation of suppressor T cells may be responsible for the development of chronic leishmaniasis.

Lack of responsiveness of beekeeper mononuclear cells to in vitro stimulation with pure bee venom

Lymphocyte proliferation activity after in vitro bee venom (BV) stimulation was examined in a group of patients allergic to bee stings and in a group of beekeepers. Although the allergic patients responded strongly to increasing doses of BV, the beekeepers demonstrated no proliferative activity and an inability to produce interleukin-2 after BV stimulation. Removal of adherent cells or various populations of suppressor cells, including Tγ cells and OKT8 positive cells, did not influence the cellular unresponsiveness of cells of beekeepers after BV stimulation. Furthermore, cells of beekeepers, when they were trypsinized or when they were preincubated for 72 hours, did not proliferate after BV challenge. It is concluded that the lack of proliferation of lymphocytes of beekeepers and the inability to produce interleukin-2 is not due to a suppressor mechanism or to the presence of anti-idiotype antibodies coating the surface of lymphocytes of beekeepers. The mechanism behind the failure of cells of beekeepers to proliferate remains unclear.

Immunohistologic Identification of Antigen-Presenting Cells in Cutaneous Sarcoidosis

Considerable evidence exists to show that activated T lymphocytes preferentially accumulate at sites of disease activity in sarcoidosis. Langerhans cells, which can be recognized by reactivity with an antibody to the T6 antigen are thought to play a primary role in T-lymphocyte activation by the skin, a tissue frequently involved in sarcoidosis. This immunohistologic study examined the distribution of OKT6-positive cells and surface expression of HLA-DR antigen in cutaneous sarcoid lesions. Skin specimens stained with an anti-HLA-DR antibody demonstrated diffuse staining of the granulomas. In addition, keratinocytes, which do not normally express HLA-DR antigens, were found to stain with monoclonal antibody to HLA-DR in an intercellular pattern. Examination of specimens for OKT6-reactive Langerhans cells revealed significantly greater concentrations in the epidermis overlying sarcoidal granulomas (33 +/- 7 cells/mm) than in the epidermis of age-, sex-, and race-matched controls (11 +/- 3 cells/mm, p less than 0.001). Of greater importance was the demonstration that significant numbers of OKT6-positive cells were present within the dermal sarcoid granulomas (19-208/mm2) in a distribution that paralleled that of Leu-3a-positive T lymphocytes. These data suggest that the epidermis may participate in activation of lymphocytes in cutaneous sarcoidosis, and implicate OKT6-positive cells in granuloma formation.

Plane Warts Under Spontaneous Regression

Immunohistologically, cellular infiltrates in regressing plane warts were mainly composed of lymphocytes and mononuclear phagocytes. There were many infiltrating T lymphocytes. Immunoelectron microscopic observation demonstrated that both helper/inducer and suppressor/cytotoxic phenotypes of T lymphocytes infiltrated in the lesions. OKT6-positive cells were observed in the dermis as well as in the epidermis. Moreover, as noted in allergic contact dermatitis, the apposition of T lymphocytes to Langerhans' cell-like cells could be seen. Lymphocytes and a small number of mononuclear phagocytes were found adjacent to damaged keratinocytes in the epidermis, the picture of which has been described as satellite cell necrosis, a hallmark of cytotoxic reaction by aggressors. These findings suggest that specific cell-mediated immunity against virus-infected keratinocytes takes place in the process of regressing plane warts.

Plane warts under spontaneous regression. Immunopathologic study on cellular constituents leading to the inflammatory reaction.

Immunohistologically, cellular infiltrates in regressing plane warts were mainly composed of lymphocytes and mononuclear phagocytes. There were many infiltrating T lymphocytes. Immunoelectron microscopic observation demonstrated that both helper/inducer and suppressor/cytotoxic phenotypes of T lymphocytes infiltrated in the lesions. OKT6-positive cells were observed in the dermis as well as in the epidermis. Moreover, as noted in allergic contact dermatitis, the apposition of T lymphocytes to Langerhans' cell-like cells could be seen. Lymphocytes and a small number of mononuclear phagocytes were found adjacent to damaged keratinocytes in the epidermis, the picture of which has been described as satellite cell necrosis, a hallmark of cytotoxic reaction by aggressors. These findings suggest that specific cell-mediated immunity against virus-infected keratinocytes takes place in the process of regressing plane warts.

Spontaneous production of eosinophil chemotactic factors by T lymphocytes from patients with subcutaneous angioblastic lymphoid hyperplasia with eosinophilia

Subcutaneous angioblastic lymphoid hyperplasia with eosinophilia (SALH) was reviewed with respect to eosinophil chemotaxis. Lymphoid cells separated from the granuloma spontaneously released at least two different eosinophil chemotactic factors (ECF): low-molecular-weight and high-molecular-weight ECF according to the profile on gel filtration (LMW-ECF, about 500; HMW-ECF, 45,000 to 70,000). The cells, however, failed to produce chemotactic activity for macrophages and neutrophils. By analysis with monoclonal antibodies against lymphocyte subpopulations, the granuloma T cells, probably OKT4-positive cells, were shown to be responsible for spontaneous production of these two ECF. Furthermore, the blood mononuclear leukocytes were separated from the patients with SALH. An ECF closely resembling HMW-ECF was also spontaneously produced by the blood OKT4-positive T lymphocytes, whereas no LMW-ECF was released. Mononuclear leukocytes from healthy donors, however, could produce an ECF resembling HMW-ECF and chemotactic activities for macrophages and neutrophils by stimulation with concanavalin A (Con A). Protein synthesis appeared to be essential for spontaneous ECF and for Con A-induced ECF production. These results suggest that the granuloma OKT4-positive T lymphocytes of the patients with SALH are in activated condition to release LMW- and HMW-ECF, whereas the blood OKT4-positive T lymphocytes are in activated condition to release only HMW-ECF. Such spontaneous and prolonged production of HMW-ECF by the cells can be one of the diagnostic means of SALH.

Disseminated pagetoid reticulosis associated with mycosis fungoides: Immunomorphologic study

We report a 77-year-old woman with erythrosquamous plaques on the legs for 10 years. Biopsies taken 4 years ago revealed a pagetoid reticulosis with a massive epidermal cell infiltrate. She has now also developed a typical tumor of mycosis fungoides. The epidermal infiltrate of the two types of lesions bore the surface membrane marker for T lymphocytes rich in both T helper and T cytotoxic-suppressor subsets (ratio, 1.8). Both lesions also showed dendritic OKT6-positive Langerhans cells, and staining of the intercellular material with the DR locus of human lymphocyte antigen was positive. A dense dermal infiltrate was evident only in the mycosis fungoides lesions, and it was similar to that in the epidermis. Electron microscopy of the two lesions showed the atypical cells described in mycosis fungoides and the Sézary syndrome. These observations suggest that disseminated pagetoid reticulosis probably is a variant of mycosis fungoides.

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.

Interleukin 2 (IL 2) up-regulates its own receptor on a subset of human unprimed peripheral blood lymphocytes and triggers their proliferation.

Several reports indicate that human peripheral blood lymphocytes (PBL) seeded in culture with purified or recombinant interleukin 2 (IL 2) immediately after separation from the blood display a substantial level of proliferation at day 5 or 6, even in the absence of any activating signal. The spontaneously IL 2 proliferating cells are large lymphocytes, and they co-purify on a Percoll gradient in the large granular lymphocytes (third (LGL) fraction) together with the natural killer (NK) activity. When LGL were separated into NKH1 (an NK-specific surface marker)-positive and NKH1-negative cells by fluorescence-activated cell sorting (FACS), proliferating cells were mainly found in the NKH1-negative fraction. On the contrary, when cells from Percoll fraction 3 were separated into OKT3-negative and positive cells, the majority of the proliferating cell was found in the OKT3-positive cells. These results indicate that spontaneously IL 2 proliferating (SIP) cells most probably belong to the T cell lineage, but are distinct from NK cells. Surprisingly, cells from this Percoll fraction examined immediately after separation from the blood do not express detectable amounts of IL 2 receptors as assessed by three different techniques: binding of [3H]IL 2, binding of [125I]anti-Tac antibodies, and FACS analysis with the use of anti-Tac antibodies. However, after 18 hr of culture in IL 2-supplemented medium, 5 to 7% of these cells became Tac-positive by FACS analysis. Additional analysis of IL 2 receptor induced in culture with IL 2 was performed by [125I]anti-TAC binding and by [3H]IL 2 binding. Scatchard analysis of [3H]IL 2 binding, in the range of concentrations leading to the detection of high-affinity binding sites, showed an affinity constant similar to that of conventional phytohemagglutinin blasts. The role of IL 2/IL 2 receptor interaction in the proliferation process was confirmed by the fact that proliferation, in contrast with NK activation, was clearly inhibited by anti-Tac antibodies. When LGL activated with IL 2 for 60 hr were sorted into Tac+ and Tac- cells, equal levels of NK activity was found in the two fractions. Proliferation, however, was only observed in the Tac+ population. We interpret these results to indicate that SIP cells are preactivated cells circulating in the blood. They are large cells and represent a very small proportion of circulating lymphocytes (0.3%). They express a subliminal amount of IL 2 receptor. Cultivated in the presence of IL 2, IL 2 receptor expression is enhanced to a detectable level, and the SIP cells begin to proliferate. These SIP cells could be activated T cells present in every normal individual.