Cells In Normal Individuals Research Articles

Changes in lymphocyte subsets in myasthenia gravis: correlation with level of antibodies to acetylcholine receptor and age of patient.

We report a detailed analysis of the subsets of lymphocytes in patients with myasthenia gravis (MG). There was a slight, nonsignificant increase in the level of CD5+ B lymphocytes among MG patients as compared with normal controls. The proportion of CD5+ T cells in MG was similar to that in controls. However, whereas age had no effect on the level of these cells in normal individuals, a significant age-related decrease of these cells was present in MG patients. The proportion of double-positive CD4+CD8+ T cells was significantly increased in MG. The level of the CD29+CD4+ (helper-inducer) subset was significantly higher in MG patients than in controls. There was no correlation between the titer of autoantibodies to acetylcholine receptor and the level of either CD29+CD4+ T cells or CD5+ B cells among MG patients. The only T-cell subset that correlated with the autoantibody titer was the CD45RA+CD4+ (suppressor-inducer) subset of CD4+ T cells.

CD4+CD7-CD57+ T cells: a new T-lymphocyte subset expanded during human immunodeficiency virus infection

CD7 and CD57 are two cell surface molecules related to the differentiation or functional stages of CD4+ T cells. The CD4+CD7- T cells represent a minor subset of CD4+ cells in normal individuals and are considered to contain the normal counterpart of Sezary T cells; the CD4+CD57+ peripheral blood lymphocytes (PBL) are detectable in long- term renal allograft recipients. We compared the cell surface expression of these CD7 and CD57 markers on CD4+ T lymphocytes in peripheral blood and lymphoid organs from normal individuals and human immunodeficiency virus (HIV)-infected patients. Our results indicate that CD4+CD7- T cells in normal PBL do not express CD57 and were poorly responsive to anti-CD3 monoclonal antibody (MoAb), the activation being restored by addition of anti-CD28 MoAb. This CD4+CD7- cell subset is increased in peripheral blood during HIV infection, and its progressive expansion mirrors both the absolute and relative decrease of CD4+ T cells. The lack of CD7 expression is correlated with CD57 acquisition on CD4+ T cells because CD4+CD7-CD57+ cells represent a major component of the CD4+CD7- subset in HIV-infected patients. Our results suggest that the presence and the expansion of CD4+CD7-CD57+ T lymphocytes, which do not behave as previously defined helper subsets, may participate to the immune dysfunction observed during HIV infection.

CD4+CD7-CD57+ T cells: a new T-lymphocyte subset expanded during human immunodeficiency virus infection

CD7 and CD57 are two cell surface molecules related to the differentiation or functional stages of CD4+ T cells. The CD4+CD7- T cells represent a minor subset of CD4+ cells in normal individuals and are considered to contain the normal counterpart of Sézary T cells; the CD4+CD57+ peripheral blood lymphocytes (PBL) are detectable in long-term renal allograft recipients. We compared the cell surface expression of these CD7 and CD57 markers on CD4+ T lymphocytes in peripheral blood and lymphoid organs from normal individuals and human immunodeficiency virus (HIV)-infected patients. Our results indicate that CD4+CD7- T cells in normal PBL do not express CD57 and were poorly responsive to anti-CD3 monoclonal antibody (MoAb), the activation being restored by addition of anti-CD28 MoAb. This CD4+CD7- cell subset is increased in peripheral blood during HIV infection, and its progressive expansion mirrors both the absolute and relative decrease of CD4+ T cells. The lack of CD7 expression is correlated with CD57 acquisition on CD4+ T cells because CD4+CD7-CD57+ cells represent a major component of the CD4+CD7- subset in HIV-infected patients. Our results suggest that the presence and the expansion of CD4+CD7-CD57+ T lymphocytes, which do not behave as previously defined helper subsets, may participate to the immune dysfunction observed during HIV infection.

Increased lymphocyte beta‐adrenergic receptor density in progressive multiple sclerosis is specific for the CD8+, CD28− suppressor cell

Beta-adrenergic receptor density on T cells from healthy humans is greatest on suppressor cells (CD8+, CD28-) and the effect of catecholamines, secreted by the sympathetic nervous system, predominates on this subset. The sympathetic skin response, a measure of sympathetic nervous system function, is absent in most patients with chronic progressive multiple sclerosis (MS). We measured beta-adrenergic receptor density on suppressor cells, cytotoxic cells, and monocytes from patients with chronic progressive MS and healthy control subjects. Control receptor density on suppressor cells was 2.8 +/- 0.3 fmol/10(6) cells versus a density of 5.1 +/- 0.7 fmol/10(6) cells for patients. Cytotoxic cell (CD8+, CD28+) receptor density was 1.4 +/- 0.4 fmol/10(6) cells in control subjects and 0.9 +/- 0.3 fmol/10(6) cells in the patients. Monocytes displayed beta-adrenergic receptor densities of 2.6 +/- 0.4 fmol/10(6) cells in normal individuals and 2.7 +/- 0.4 fmol/10(6) cells in the patient group. CD8 lymphocyte beta-adrenergic receptor densities in patients with relapsing-remitting and those with stable MS were not different from control values, yet were significantly less than the values for patients with chronic progressive MS. We find that mononuclear cells from healthy control subjects and patients with chronic progressive MS proliferate in response to 200 units/ml of recombinant human interleukin-2 (IL-2) similarly. However, IL-2 treatment increased beta-adrenergic receptor density on normal mononuclear cells, but failed to increase it on mononuclear cells from patients with chronic progressive MS.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of testicular autoantigens and influence of lymphokines in testicular autoimmune disease

The present data on testicular autoimmune disease suggest that CD 4+ helper type T cells are responsible for adoptive transfer of murine experimental autoimmune orchitis (EAO). In passive EAO, the earliest lymphoid cell infiltration is confined to terminal segments of seminiferous tubules. Both orchitis and vasitis develop. In active EAO, any location in the testis is affected, frequently involving subcapsular seminiferous tubules. The location of maximum histopathology in passive EAO coincides with the site in the normal testis that expresses maximal Ia. Demonstration of IgG in the testis after immunization with testicular antigens or after transfer of sera from orchiectomized mice immunized with testis shows that autoantigens are present on the germ cells outside the Sertoli cell barrier, suggesting the existence of dynamic protective mechanisms against immune responses to the non-sequestered autoimmunogenic germ cells in normal individuals.

Flow cytometric analysis of erythrocyte populations in tn syndrome blood using monoclonal antibodies to glycophorin A and the Tn antigen

Flow cytometric analysis employing monoclonal antibodies to the Tn antigen and glycophorin A was used to characterize the erythrocyte populations present in blood samples from individuals with Tn syndrome. Four monoclonal antibodies specific for the Tn antigen, Gal-NAc monosaccharide, on human erythrocytes were obtained from a fusion of splenocytes from a Biozzi mouse immunized with red cells from a Tn individual. These monoclonal antibodies specifically recognize GalNAc monosaccharide sites located on the erythrocyte cell surface sialoglycoproteins, glycophorin A and glycophorin B, and do not bind to fixed normal red cells presenting the Neu-NAc alpha 2-3Gal beta 1-3(NeuNAc alpha 2-6)GalNAc alpha 1-O-Ser(Thr) tetrasaccharide or to fixed neuraminidase-digested cells presenting the Gal-GalNAc disaccharide. The percentages of Tn-positive red cells in samples from six unrelated Tn donors ranged from 28 to 99%. Binding of the glycophorin A-specific monoclonal antibodies showed that the erythrocytes composing the Tn-negative fraction presented normal amounts of the M and N epitopes on glycophorin A. The presumed somatic mutational origin of Tn-positive cells was tested in blood samples from five normal donors; three possible Tn cells were observed after analysis of a total of 1.1 x 10(7) erythrocytes, suggesting that the frequency of such cells in normal individuals is less than 1 x 10(-6).

Autoimmunity and the pathogenesis of glomerulonephritis

Self-tolerance is maintained by: thymic influences on developing T cells; peripheral mechanisms that can tolerise post-thymic T cells; and to a variable extent the tolerisation of potentially autoreactive B cells. The presence of autoreactive T cells in normal individuals suggests that mechanisms to control the activity of such cells may be important. Failure of any of these processes may lead to autoimmunity. The relationship between glomerulonephritis and the mechanisms leading to breakdown of self-tolerance remains elusive. An important observation is that autoimmune diseases are strongly associated with particular products of the major histocompatibility complex (MHC). This association may reflect the intimate involvement of the MHC in thymic T cell education. Another explanation is that T cells only recognise antigens presented in the context of MHC molecules. Although there has been progress in identifying the targets recognised by autoantibodies in vasculitis and anti-GBM disease, nothing is known about the T cells involved. Despite our ignorance, therapy aimed specifically at the T cell can be effective. This approach is being supplemented by attempts to engage immunoregulatory mechanisms, such as idiotype-antiidiotype interactions. The hope is that such treatments, or combinations thereof, will allow a more focused suppression of the autoimmune response, in contrast to the non-specific (and therefore potentially dangerous) methods of immunosuppression available at present.

Detection of Human Papillomaviruses in Cervicovaginal Cells Using Polymerase Chain Reaction

The polymerase chain reaction (PCR) was used to identify human papillomavirus (HPV) in cervicovaginal cells in normal individuals and in patients with cervical intraepithelial neoplasia (CIN). By use of a set of primers encoding the E6 region of the HPV genome, the presence of HPV DNA was demonstrated in the cervicovaginal cells of 43 (42.2%) of 102 normal individuals and in all 12 CIN patients. High sensitivity of the PCR method produced an additional 9 positive results on second sampling from 48 individuals who were initially HPV-negative. On the other hand, 26 (24.3%) of 107 HPV-positive individuals were HPV-negative when sampled a second time 5-7 days later. The data suggested that retest probably should be considered for patients clinically suspected of having HPV infection whose initial test results are negative for HPV DNA. Also, single HPV DNA-positive results should be accepted with caution.

HIV infection is associated with the spontaneous production off interleukin-1 (IL-1) in vivo and with an abnormal release off IL-1αaL in vitro

Interleukin-1 (IL-1) is not constitutively produced by normal human monocytes. We have investigated the production of cell-associated IL-1 in uncultured unstimulated adherent monocytes from HIV-infected patients, which reflects ongoing generation of IL-1 by the cells in vivo. High levels of cell-associated IL-1 activity and of cell-associated IL-1 alpha and IL-1 beta antigens were found in monocytes from HIV-infected patients as compared with those found in monocytes from normal individuals. Amounts of cell-associated IL-1 were high in patients with AIDS and in patients from Centers for Disease Control groups II and III. Serum-free culture for 24 h of monocytes from HIV-infected individuals in the absence of lipopolysaccharides (LPS) resulted in spontaneous release of IL-1 activity from the cells whereas no release occurred upon culture of normal cells. Stimulation of monocytes with LPS induced the release of IL-1 alpha and IL-1 beta from cells of infected patients. Only IL-1 beta was released from cells of normal individuals. Thus, circulating monocytes from HIV-infected patients are triggered to produce IL-1 in vivo. The present study also indicates that HIV infection is associated with an acquired defect in the intracellular processes regulating IL-1 secretion.

Characterization of a T-cell subset prevalent in immunoregulatory disorders in humans

T cells from individuals with certain autoimmune diseases (rheumatoid arthritis, graft-versus-host disease, acquired immunodeficiency syndrome) express high levels of a cell surface sialoglycoprotein with a molecular weight of 140 kDa (gp140). Although a low frequency of gp140 + T cells was detected in the blood of normal individuals, upon stimulation with autologous EBV-transformed B cells (AMLR), the frequency of expression of gp140 was increased threefold. To further characterize gp140 + T cells, rosetting techniques with ox erythrocytes coated with monoclonal anti-gp140 antibody were used to isolate T-cell subsets for phenotypic, cell cycle, and functional analysis. The majority of gp140 + T cells expressed cytotoxic/suppressor (CD8 +) phenotype in both normal and AMLR-activated states. Unstimulated gp140 + T cells had significantly greater nucleic acid content, as measured by acridine orange and flow cytometry, than gp140 − T cells. Surprisingly, the gp140 + T-cell subset had a less proliferative response in vitro to pokeweed or phytohemaglutinin mitogens. These results suggest that gp140 + T cells in normal individuals and in patients with autoimmune diseases may have been activated previously in vivo and that they are relatively resistant to reactivation in vitro.

Phenotypic and functional deficiency of natural killer cells in patients with chronic fatigue syndrome.

Natural killer (NK)3 cells are large granular lymphocytes that appear to play a significant role in the host's defense against viral infection. We performed an extensive phenotypic and functional characterization of NK cells on 41 patients with the chronic fatigue syndrome (CFS), or "chronic active Epstein-Barr virus infection" syndrome, and on 23 age- and sex-matched asymptomatic control subjects in an attempt to further characterize this illness. These studies demonstrated that a majority of patients with CFS have low numbers of NKH1+T3- lymphocytes, a population that represents the great majority of NK cells in normal individuals. CFS patients had normal numbers of NKH1+T3+ lymphocytes, a population that represents a relatively small fraction of NK cells in normal individuals. When tested for cytotoxicity against a variety of different target cells, patients with CFS consistently demonstrated low levels of killing. After activation of cytolytic activity with recombinant interleukin 2, patients were able to display increased killing against K562 but most patients remained unable to lyse Epstein-Barr virus-infected B cell targets. Additional cytotoxicity experiments were carried out utilizing anti-T3 monoclonal antibody to block killing by NKH1+T3+ cells. These experiments indicated that the NK cell that appears to be responsible for much of the functional activity remaining in patients with CFS belongs to the NKH1+T3+ subset, which under normal circumstances represents only approximately 20% of the NK cell population.

Histamine levels in bronchoalveolar lavage from patients with asthma, sarcoidosis, and idiopathic pulmonary fibrosis

Bronchoalveolar lavage (BAL) has been used extensively as a research tool to elucidate immunologic events occurring in the lower respiratory tract of patients with numerous diseases and, most recently, to study patients with asthma. We assessed mast-basophiloid cell numbers and histamine levels with a sensitive histamine assay, lower limit of sensitivity, 25 pg/ml, in BAL fluid from normal individuals (n = 9) and compared these results to those obtained from patients with sarcoidosis (n = 31), idiopathic pulmonary fibrosis (IPF) (n = 8), and mild asthma (n = 7). Patients with sarcoidosis demonstrated a significant increase in total BAL mast-basophiloid cells, 9.6 ± 4.1 × 10 4, compared to total cells in normal individuals, 0.0, p = 0.03, whereas only patients with IPF had significant elevations in BAL histamine levels. 1315 ± 737 pg/ml, versus levels in normal individuals, 161 ± 54 pg/ml, p = 0.002. A good correlation existed between histamine levels on an aliquot of lysed BAL cells and BAL histamine levels, R = 0.655 and p = 0.02, but not with either the total number or percent mast-basophiloid cells in BAL assessed on Wright's stained cytocentrifuge preparations. Subjects with asthma had both normal numbers of BAL mast-basophiloid cells and histamine levels. These data suggest that (1) BAL histamine levels are easily quantified, (2) the reason(s) for elevations in BAL histamine levels in IPF need further investigation, (3) BAL histamine levels in subjects with asthma are not elevated in those with mild and stable disease, and (4) lumenal mast-basophiloid cells are one major source of BAL histamine.

Thyroid autoimmunity in patients on long term therapy with leukocyte-derived interferon.

Among 49 patients with carcinoid tumors given long term therapy (mean, 8 months; range, 3-36) with human leukocyte-derived interferon-alpha (huLe-IFN alpha), hypothyroidism occurred in 5 and thyrotoxicosis in 2. Antibodies against thyroid microsomal antigen and/or thyroglobulin were found in 13 patients. In 7 of these, 3 of whom developed hypothyroidism, the antibodies appeared after the start of therapy. During treatment, an increase in the proportions of circulating activated surface HLA-DR-positive T-helper and T-suppressor cells occurred after 3-4 days, and the proportions remained elevated at 3 and 6 months. Incubation of T cells of normal individuals in vitro with the huLe-IFN alpha preparation induced a rise in activated T-helper and T-suppressor cells. This effect was mimicked by recombinant IFN gamma (r-IFN gamma), but not by r-IFN alpha. Further, the huLe-IFN alpha preparation employed induced HLA-DR expression on human thyroid cells in tissue culture as did r-IFN gamma, but not r-IFN alpha, suggesting the presence of bioactive IFN gamma in the huLe-IFN alpha preparation. The results demonstrate that thyroid autoimmune disease can occur as a side-effect of treatment with huLe-IFN alpha and suggest that IFNs may play important regulatory roles, at both the effector and target cell levels, in the development of human autoimmune disorders.

Immunomodulation by Bronchial Lavage Cells in Normal Individuals and Patients with Bronchogenic Carcinoma

The ability of bronchoalveolar lavage cells to facilitate lymphoproliferation to mitogen in a system which allows assessment of pulmonary alveolar macrophage accessory cell function was investigated. Bronchoalveolar lavage cells were obtained from healthy non-smokers and smokers and from patients undergoing diagnostic bronchoscopy. Lavaged cells were cultured with monocyte-depleted homologous blood lymphocytes obtained from healthy, young volunteers and stimulated with suboptimal (2 micrograms/ml) or optimal (20 micrograms/ml) concentrations of phytohemagglutinin. Mitogen responses of lymphocytes in all groups were related to the number of lavage cells added, increasing with 1:100 and 1:10 bronchoalveolar lavage cell to lymphocyte ratios and decreasing with 1:2 and 1:1 ratios. Lymphoproliferative responses observed in smoker and nonsmoker cultures were not different. In contrast, maximal proliferative responses of cell cultures from patients with epidermoid and small cell carcinoma were decreased compared with cultures from patients with adenocarcinoma or controls. These data show that pulmonary bronchial lavage cells from smokers and nonsmokers provide similar dose related augmentation and suppression of lymphocyte mitogenic responses. Furthermore, accessory cell function of lavage cell populations is normal in patients with adenocarcinoma, but depressed in patients with epidermoid or small cell carcinoma.

T-cell reactivity to polymorphic MHC determinants. II. Self-reactive and self-restricted T cells.

Alloreactive T cells recognize polymorphic nonself MHC determinants that are expressed by different members of the same species. They are detected in specifically stimulated cytotoxic/proliferative responses in vitro (allogeneic mixed leukocyte reaction, allo MLR), and in the graft-versus-host (GvH) or host-versus-graft (HvG) reactions of various transplantation systems in vivo. These primary cell-mediated immune responses are easier to induce and are apparently of a different nature than MHC-restricted T-cell-mediated immune responses to conventional antigens or pathogens. This has always intrigued immunologists (and frustrated transplantation activists). Attempts to explain this preponderance of the T-cell reactivity to allogeneic MHC determinants focused either on a particular “antigenic nature” of MHC-encoded glycoproteins, or assumed a large pool of precursor T cells in normal individuals that specifically recognized allogeneic MHC antigens.

T-cell reactivity to polymorphic MHC determinants. III. Alloreactive and allorestricted T cells.

Alloreactive T cells recognize polymorphic nonself MHC determinants that are expressed by different members of the same species. They are detected in specifically stimulated cytotoxic/proliferative responses in vitro (allogeneic mixed leukocyte reaction, allo MLR), and in the graft-versus-host (GvH) or host-versus-graft (HvG) reactions of various transplantation systems in vivo. These primary cell-mediated immune responses are easier to induce and are apparently of a different nature than MHC-restricted T-cell-mediated immune responses to conventional antigens or pathogens. This has always intrigued immunologists (and frustrated transplantation activists). Attempts to explain this preponderance of the T-cell reactivity to allogeneic MHC determinants focused either on a particular “antigenic nature” of MHC-encoded glycoproteins, or assumed a large pool of precursor T cells in normal individuals that specifically recognized allogeneic MHC antigens.KeywordsLymphoblastoid Cell LineResponder CellLimit Dilution AnalysisReceptor PhenotypeLimit Dilution CultureThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Kinetic studies on lymphocytes labelled with indium 111‐oxine in patients with chronic lymphocytic leukaemia

The kinetics of 111Indium-oxine-labelled autologous blood lymphocytes were studied in normal subjects and 10 patients with chronic lymphocytic leukaemia (CLL). In both normal and leukaemic subjects, intravascular survival of lymphocytes was characterized by two exponential decreases, the first being a rapid one and the second slow phase. T 1/2 of the second phase for normal T lymphocytes (52.0 +/- 3.2 h: mean +/- SEM) was longer than that for B lymphocytes (31.6 +/- 2.8 h). T 1/2 of the second phase in patients with CLL was 15.1 to 192.6 h, which correlated well with the clinical stage of CLL by the Rai classification. Body surface scanning revealed prominent splenic accumulation of labelled cells in normal individuals and patients with CLL. Lymph node visualization was recognized in all patients with T-cell CLL; however, there was no visualization in 8 of the 9 patients with B-cell CLL. The longer survival time and splenic and lymph node visualization suggest that an expansion of the extravascular lymphocyte pool may be important in the pathophysiology of CLL.

Peripheral blood lymphocytes from normal individuals can be induced to secrete immunoglobulin G antibodies against self-antigen thyroglobulin in vitro.

Autoantibodies to the self-antigen thyroglobulin (Tg) are usually not found in sera of normal individuals, but are often present in sera of patients with autoimmune thyroid diseases. To determine if the presence of such autoantibodies could be due to the abnormal appearance of self-reactive B cells, which are otherwise absent in normal subjects, or to an alteration in the mechanisms regulating such B cells, peripheral blood lymphocytes (PBL) from normal individuals and patients with autoimmune thyroid diseases were cultured and stimulated in vitro with the polyclonal stimulant pokeweed mitogen (PWM). A modified plaque assay was used to enumerate cells secreting protein A-binding immunoglobulins (Igs) and specific antibodies against Tg. PBL from all individuals tested, including normal subjects (n = 26), could be induced by PWM to produce antibodies against Tg in vitro and these antibodies were of IgG isotypes. PBL from patients with detectable serum anti-Tg had more inducible cells secreting anti-Tg [27,000 +/- 10,700 (+/- SD)/10(6) PBL] than those from patients with autoimmune thyroid diseases, who had no detectable serum anti-Tg (8,000 +/- 5,000), and those from normal individuals (7,200 +/- 4,200). The demonstration of inducible mature (IgG) anti-Tg-producing cells in normal individuals suggests that subclinical autoimmunity against certain self-antigens may be a normal phenomenon in man and that its escalation into clinical autoimmune conditions is prevented through regulation of the specific self-reactive cells.

Studies on the suppression of lymphocyte proliferation by concanavalin-A activated mononuclear cells in normal individuals.

Studies on the suppression of lymphocyte proliferation by concanavalin-A activated mononuclear cells in normal individuals.

Analysis of Natural Killer Cell Cytotoxicity of Cancer Patients Treated With Recombinant lnterferon<xref ref-type="fn" rid="FN2">2</xref><xref ref-type="fn" rid="FN3">3</xref>

Peripheral blood natural killer (NK) cell cytotoxicity of 24 cancer patients was studied prior to and after single and multiple injections of various doses of human leukocyte recombinant interferon-alpha clone A (IFN-alpha rA). The NK cell cytotoxicity of all cancer patients declined consistently 4 and 8 hours after a single injection of IFN-alpha rA. Twenty-four hours after the injection of IFN-alpha rA, NK cell cytotoxicity of patients with low NK cell phenotype (NK-LR) was significantly augmented, whereas that of patients with medium (NK-MR) or high (NK-HR) NK phenotype was depressed. After multiple injections of IFN-alpha rA, depression of NK cell cytotoxicity was observed in a number of NK-MR and NK-HR patients, but in some patients with NK-LR phenotype, further potentiation was observed. No direct correlation between the NK cell augmentation and serum IFN levels was detected. In in vitro studies, IFN-alpha rA, when added to cultures of target and effector cells of normal individuals in a dose of 10(3) U/ml, was efficient in augmenting NK cell cytotoxicity. NK cell cytotoxicity of cancer patients could also be augmented by the IFN-alpha rA preparation; however, this augmentation occurred only prior to in vivo IFN-alpha rA therapy. After IFN-alpha rA in vivo therapy, their NK cells became refractory to further in vitro IFN-alpha rA treatment.