Tuberculous anergic sera or purified protein derivative treatment induces modification in lymphocyte transformation of cells from patients with tuberculosis.

Peripheral blood mononuclear cells from 20 purified protein derivative (PPD)-reactive (PPD+) tuberculous patients were cultured in autologous, tuberculous anergic, or normal serum. After 12 h of incubation, the serum was eliminated and lymphocyte transformation with PPD was performed. Transformation was inhibited only in cells incubated with anergic serum. In contrast, cells from 11 anergic tuberculous (PPD-) individuals recovered the ability to respond to an optimal PPD dose after treatment with high PPD concentrations followed by several washings. The cells which recovered returned to their initial anergic state when incubated with sera from anergic patients. Under both conditions, incubation with sera did not abolish the response to the mitogen phytohemagglutinin. Cells from healthy PPD+ or PPD- individuals were used as controls. The most important finding derived from serum analysis was the increased levels of specific immunoglobulins G and A in anergic patients.     

Augmentation of guinea pig T lymphocyte proliferative response to antigens in the presence of purified B cells

The effect of supplemental B lymphocytes on the antigen-induced in vitro proliferative response of highly purified lymph node T cells was studied in two immune systems of complete Freund's adjuvant (CFA) and testicular antigen (TA) sensitized guinea pigs. When the T lymphocytes obtained from animals 2 weeks after CFA sensitization, though unresponsive to purified protein derivate of tuberculin (PPD) by themselves, were cultured together with mitomycin C-treated autologous B (Bm) lymphocytes (1:0.5 ratio), they responded well to PPD. The enhancing effect of Bm lymphocytes was less than that of mitomycin C-treated macrophages (M phi m) from the same animals. Analogous helper effect of Bm lymphocytes on the T cell proliferation was observed also in TA-sensitized animals. Additionally, it is noteworthy that the PPD-induced proliferation of T lymphocytes at 2 weeks after CFA sensitization was absolutely dependent on M phi and B lymphocytes, whereas T lymphocytes at 10--14 weeks exhibited a low but significant response by themselves, although their response to PPD was markedly augmented with supplemental M phi m or Bm lymphocytes.     

TG and TnonG lymphocyte involvement in the proliferative response to PHA. I. Reactivity and regulatory functions of TG and TnonG subsets in the proliferative response of T lymphocytes

Both TG and TnonG subpopulations from human peripheral blood were found to secrete helper and suppressor factors for the PHA-induced T cell proliferative response. The TnonG subpopulation contained cells which were both able to secrete helper factor and to respond by proliferation, whereas in the TG subpopulation cells with a proliferative potential could not be conclusively demonstrated. Suppressor and helper activities in TG and TnonG culture supernatants displayed a different time kinetics and were influenced by the dose of mitogen in test cultures.     

Low lymphocyte interferon-gamma production and variable proliferative response in anorexia nervosa patients

Interferon-gamma (IFN-) production by peripheral blood mononuclear cells (PBMC) in 14 patients with anorexia nervosa (AN) was significantly lower than in 14 age-matched healthy controls. Follow-up samples in four patients displayed low levels, except in two when they recovered the IFN- production as the hormonal cycles were restored. A large interindividual variation for the lymphocyte proliferative response was observed in 30 AN patients. DNA synthesis of PBMC was normal in 8 patients (27%), significantly increased in 6 (20%) (P<0.001), and significantly decreased in 16 (53%) (P<0.001). IFN- inhibition was reversed by culturing a control lymphocyte population with monocytes from patients with AN. This was not observed in cultures of control monocytes and AN lymphocytes. IL-2 receptor (TAC subunit) was assessed and no difference was found in the number of TAC-positive cells between patients and controls. These results point out impaired production of the immunomodulator cytokine IFN- as a major functional defect of AN peripheral lymphocytes.     

Enhancement of human lymphocyte proliferative response to purified protein derivative by an anti-interleukin-2 receptor alpha chain antibody (CD25).

While it is clear that the beta subunit of interleukin-2 receptor (IL-2R) plays a pivotal role in IL-2-induced signal transduction, the function of the alpha subunit, other than modulating the association rate of IL-2, is still unknown. It has been reported that the interaction between IL-2 and the IL-2R alpha subunit of several IL-2-dependent murine T-cell lines may result in a negative regulatory signal. To confirm this finding, we investigated the effect of an anti-IL-2R alpha antibody, CD25-8D8, on the proliferative response of human peripheral blood lymphocytes. Lymphocytes from purified protein derivative (PPD)-positive donors were cultured with PPD and various concentrations of CD25-8D8 for up to 9 days, and [3H]thymidine uptake was measured. Whereas the proliferative response of human lymphocytes to PPD was suppressed by high concentrations of CD25-8D8, subinhibitory amounts of CD25-8D8 enhanced lymphocyte proliferation by 3.5-fold (range 2.2-6.2-fold) on the second day after maximal [3H]thymidine uptake had occurred. By itself, CD25-8D8 could not induce proliferation of washed 5-day PPD-activated lymphocytes during reculturing; instead, growth enhancement by CD25-8D8 was dependent on the presence of PPD-activated culture supernatant or moderate levels of exogenous IL-2. The enhancing effect of anti-IL-2R alpha antibody, observed in both murine and human systems, reinforces the possibility that binding of IL-2 to the IL-2R alpha chain plays a negative regulatory role in signal transduction.     

T-dependence of human B lymphocyte proliferative response to mitogens.

Human peripheral blood and tonsil lymphocytes were fractionated on anti-Ig-coated Sephadex columns or by centrifugation after rosetting with native sheep erythrocytes. Both methods allowed the recovery of B and T-enriched populations the purity of which was checked by fluorescein-labelled anti-Ig serum, E and EAC rosette formation, and heterologous antisera specific for B or T lymphocytes. The proliferative response of T cells to PHA, Con A, PWM, and ALS was not found different from that of unfractionated cells, whereas no response of the B cells could be observed to these mitogens providing that no contaminating T cells were present. Addition of T lymphocytes to these unresponsive B cells allowed them to respond to phytomitogens, but not to ALS. X-irradiated T cells could, to some extent, replace the diving T lymphocytes; no T-replacing factor could be found in cell-free supernatants from T cells, whether or not they had been activated by mitrogens. This model of B-T cooperation appears useful for studying the differentiation and maturation of human B lymphocytes.     

Adrenergic regulation of lymphocyte proliferative response in cultures with T-cell mitogens

The effects of epinephrine, β-adrenergic agonist terbutaline sulfate, and cAMP phosphodiesterase inhibitor theophylline on proliferative response of peripheral blood lymphocytes from healthy subjects were studied in cultures with phytohemagglutinin and concanavalin A. Both adrenergic agonists inhibited lymphocyte blastogenesis, but the effect of epinephrine was more pronounced. Translated fromByulleten' Eksperimental' noi Biologii i Meditsiny, Vol. 128, No. 8, pp. 207–209, August, 1999     

Prostaglandin-dependent regulation of the in vitro proliferative response to mycobacterial antigens of peripheral blood lymphocytes from normal donors and from patients with tuberculosis or leprosy.

The effect of age on the in vitro generation of immunoglobulin-secreting cells in pokeweed mitogen-stimulated cultures was examined using a staphylococcal protein A plaque assay. Although there was no statistically significant decrease with age in the numbers of plaque-forming cells, subjects whose cells failed to produce immunoglobulin were four times more common amongst individuals over 55 years of age. Simultaneously-measured T and B lymphocyte numbers. 3H-thymidine incorporation by mitogen-stimulated cultures, and serum immunoglobulins were comparable in both the young and the aged.     

TG and TnonG lymphocyte involvement in the proliferative response to PHA. II. The role of TG and TnonG lymphocytes in the regulation of the PHA-induced proliferative response of non-T cells

The TnonG subpopulation from human peripheral blood provides help for PHA induced proliferation of non-T cells, while the TG subpopulation does not. Thus the inducer cells in the proliferative response of T and non-T cells to PHA are at least in part different.     

Subpopulation structure of T-regulatory cells and proliferative response of blood lymphocytes in vitro under pulmonary tuberculosis

Subpopulation structure of T-regulatory cells and proliferative activity of blood lymphocytes in vitro in patients with different clinical forms of pulmonary tuberculosis were studied in this work. It has been shown that Trn--natural T-regulatory lymphocytes (CD4+CD25+Foxp3+) play a leading role in formation of immune suppression under infiltrative, disseminated and fibrosis-cavernous pulmonary tuberculosis. Besides, their number is increased in blood of both tuberculin-positive and tuberculin-negative patients. Negative correlation between the number of Trn and proliferative activity of blood lymphocytes (basal, mitogen- and antigen-induced)has been established, which testifies about participation of Trn in suppression of lymphocyte proliferation and Th1- and Th2-immune response.     

T and B lymphocytes in patients with pulmonary tuberculosis.

Counts of T and B lymphocytes were made in the peripheral blood from patients with active and inactive pulmonary tuberculosis and in healthy subjects. In patients with active pulmonary tuberculosis the numbers of T cells were significantly lower than in the controls. After treatment for two months, if the patient's condition improved the percentage of T cells returned to nearly normal levels. If the patient failed to improve, the numbers of T cells remained low. The mechanism of these phenomena is discussed.     

Peripheral blood lymphocyte subpopulations in patients with primary proliferative and secondary polycythaemia.

Peripheral blood lymphocyte subpopulations were measured in 18 patients with primary proliferative polycythaemia and 13 patients with secondary polycythaemia. A decrease in numbers of suppressor T lymphocytes and an increase in the helper:suppressor T lymphocyte ratio was found in those with primary polycythaemia compared with normal subjects and patients with secondary polycythaemia. If other causes of an increased helper:suppressor ratio are excluded this variable may be useful in confirming the myeloproliferative nature of patients with erythrocytosis.     

T-cell proliferative response to antigens secreted by Mycobacterium tuberculosis.

An infection model of human tuberculosis was established with C57BL/6J mice. The lymphocyte proliferative responses to antigens from Mycobacterium tuberculosis were investigated during the course of infection and compared with results obtained with a group of mice immunized with large amounts of killed bacteria. The two groups responded similarly to a number of mycobacterial antigens, but marked differences in responses against secreted antigens were found; only infected mice responded vigorously to these. The responding lymphocyte subpopulation was made up of L3T4+ T lymphocytes under restriction of the Ia molecule.     

In vitro differences in lymphocyte subpopulation reactivity in lung cancer patients: purified protein derivative-specific suppressor T lymphocytes in patients who have received Bacillus Calmette-Guerin

Antigen-specific responses of lymphocyte to purified protein derivative (PPD) were studied in seven patients with non-small cell lung cancer who had undergone complete resection and received postoperative intrapleural Bacillus Calmette-Guerin. In addition, one patient with unresected lung cancer and reactive tuberculin skin test (PPD +) and five normal PPD- individuals were also studied. Lung cancer patients had significantly fewer responsive peripheral blood lymphocytes, unfractionated T cells, and T cells bearing Fc-IgG receptors (TG + populations) than normal controls. These deficits were most pronounced in the group who had received intrapleural Bacillus Calmette-Guerin but who had failed to develop reactive tuberculin skin tests. In contrast, the TG-populations (FC-IgG receptor-negative T cells) from all patients responded to PPD. TG + cells specifically inhibited TG- -cell responses to PPD in both proliferation and immunoglobulin secretion. Radiosensitive suppressor monocytes were found in other patients. This study shows interesting immune deficits in early lung cancer patients. These patients appear to have PPD-specific suppressor TG + cells which may contribute to the immune deficits in these patients.     

Tuberculin purified protein derivative-reactive T cells in cord blood lymphocytes.

Lymphocytes obtained from cord blood of newborn babies who were born of healthy mothers were studied in vitro for their responsiveness to purified protein derivative (PPD) of tuberculin. Cord blood lymphocytes proliferated in vitro by stimulation with PPD, despite wide variations in the results. Studies with fractionated lymphocytes revealed that PPD-responding cells belonged to E-rosetting, nylon wool-nonadherent T lymphocytes. Non-E-rosetting B lymphocytes alone did not proliferate at all after stimulation with PPD. In addition, bromodeoxyuridine and light treatment of in vitro PPD-stimulated lymphocytes eliminated the responsiveness to PPD. These results suggest that T lymphocytes do exist in cord blood and respond in vitro to stimulation with PPD. A possible role for PPD-reactive T lymphocytes in cell-mediated protective immunity in newborns is discussed.     

Activated mouse lymphocyte release factors which modulate the proliferative responses of other lymphocytes.

Supernatants of mouse lymphocyte cultures briefly exposed to phytomitogens or various antigens may contain factors---lymphokines--which are mitogenic for lymphocytes in the absence of any additional stimulatory agents. Whether such supernatants can modulate the proliferative responses of lymphocytes exposed to phytomitogens or allogeneic cells is the subject of the present investigation. Active supernatants were obtained from cultures of either mouse lymph node cells incubated with concanavalin A bound to Sepharose or allogeneic cells for 24 h. Both types of active supernatants reduced the 3H-thymidine uptakes of lymph node cells and cortisone-resistant--medullary--thymocytes in response to various phytomitogens and allogeneic cells. In contrast, normal suspensions of mouse thymocytes, containing both cortical and medullary lymphocytes, exhibited stimulations higher than anticipated. The results indicate that activated lymphocytes may release factors which reduce proliferative responses of some lymphocytes, presumably immunologically mature T cells, and enhance the responses of other lymphocytes which are relatively immature.     

Nonspecific stimulation of lymphocytes by tuberculin.

Lymphocytes from uninfected, nonimmune mice can be stimulated to proliferate and produce antibody to diverse antigens in culture when exposed to the purified protein derivative of tuberculin or unheated tuberculin culture filtrate. Lymphocytes from numerous inbred strains of mice respond to tuberculin, but no true low responder strain has been found as yet. The evidence presented suggests that a subpopulation of B cells, which are distinct from those activated by lipopolysaccharide endotoxin, respond to tuberculin. The results of digestion experiments with insolubilized enzymes suggest that the nonspecific activating principles in tuberculin are protein.     

Migration of blood-borne lymphocytes into antigen-stimulated lymph nodes. A study of the kinetics of lymphocyte recruitment throughout an immune response

The kinetics of lymphocyte recruitment into rat popliteal lymph nodes stimulated with varying doses of allogeneic lymphocytes or purified protein derivative of tuberculin differed markedly. The number of blood-borne lymphocytes entering lymph nodes stimulated with allogeneic lymphocytes was greater and the duration of their entry more prolonged than in lymph nodes stimulated with the soluble antigen purified protein derivative of tuberculin. When individual antigenic doses were compared, lymphocyte recruitment into popliteal lymph nodes was shown to be dependent upon antigenic dose; higher doses of antigen resulting in increased levels of lymphocyte recruitment. In addition it was found that there is no linear relationship between lymph node weight changes and the extent of lymphocyte recruitment.