Spontaneous Proliferative Response Research Articles

Effectiveness of allergen-specific immunotherapy in children with bronchial asthma sensitized to house dust mites

Background. The only method of treatment that can change the natural course of allergies is allergen-specific immunotherapy, which allows reducing the sensitivity of the child’s body to the causative allergen and decreasing the activity of chronic inflammation. The evidence base for the effectiveness of allergen-specific immunotherapy in the treatment of bronchial asthma (BA) has significantly expanded recently. However, the truly important results remain too limited to make clinically useful conclusions about the efficacy of sublingual allergen-specific immunotherapy (SLIT) in people with BA and further evidence is needed. The purpose of the study is to evaluate the effectiveness of SLIT as an additional treatment for atopic BA of varying severity in children sensitized to house dust mites. Materials and methods. The main group consisted of 80 children with BA aged from 5 to 17 years, they were divided into three groups according to the severity of the disease: group 1 — 20 children with mild intermittent asthma, group 2 — 30 children with mild persistent asthma, group 3 — 30 children with moderately severe persistent asthma. Thirty practically healthy children without BA were also examined as a control group. For SLIT, a sublingual ORALTEK spray produced by Inmunotek (Spain) was used, a mixture of house dust mite allergens Dermatophagoides pteronyssinus 50 %/Dermatophagoides farinae 50 %, 1 ml of the solution contains 30,000 IU of natural allergen extract. To assess the effectiveness of the treatment, we conducted an analysis of the total score of clinical manifestations of asthma and allergic rhinitis and the degree of asthma control according to the Asthma Control Test. We also analyzed the state of cellular and humoral immunity in the dynamics of therapy. These indicators were evaluated within 1 month before the beginning and a month after the end of the one-year course of therapy. To assess the validity and reliability of the obtained results, statistical data processing was carried out using Microsoft Excel 2016 and Statistica for Windows 10 programs. The Student’s t-test was applied to assess the reliability of the results. The difference between the compared values was considered statistically significant at p < 0.05. Results. The results of the statistical analysis confirmed a significant improvement in the control indicators of patients who received SLIT: reduction in the total score of clinical manifestations, the frequency of clinical symptoms and the use of emergency drugs (p < 0.05). By the end of the study, the dynamics of a decrease in these indicators compared to the data before the start of treatment (according to the above list) was as follows: in patients of group 1 — 82.5, 91.8, 94.5 %, in patients of group 2 — 80.5, 88.4, 92.7 %, in patients of group 3 — 76.7, 85.3, 91.2 %. After treatment with the addition of SLIT to the basic therapy, a reliable increase in IgE and sIgA was observed, an increase in the spontaneous proliferative response of T-lymphocytes to the mitogen in all three groups to the level of the controls, a probable decrease in eosinophilic inflammation and an improvement in indicators of the cellular and humoral immunity (p < 0.001). Conclusions. The prescription of SLIT to children with mild intermittent BA, mild persistent BA and perennial allergic rhinitis, moderate persistent BA and perennial allergic rhinitis provides an opportunity to obtain stable control of the disease, reduces the number of visits to the doctor and justifies the wide use of this method in clinical allergy practice.

T cell receptor repertoire and mitotic responses of lamina propria T lymphocytes in inflammatory bowel disease.

Human intestinal lamina propria T lymphocytes (LPL-T) physiologically exhibit minimal proliferation in response to antigen receptor stimulation in vitro. This is thought to occur as a consequence of regulatory influences which are exerted by the mucosal microenvironment. The present study is aimed at investigating whether proliferative responses of intestinal LPL-T to antigen receptor stimulation are altered in patients with inflammatory bowel disease. Accordingly, proliferative responses of LPL-T in patients with Crohn's disease and ulcerative colitis to stimulation with CD3 MoAb plus IL-2 were examined and compared with controls. In addition, T cell receptor (TCR) repertoires of LPL-T and peripheral blood T lymphocytes were determined by indirect immunofluorescence using a panel of 11 TCR V beta specific antibodies. In most patients with inflammatory bowel disease, LPL-T showed enhanced proliferation to antigen receptor stimulation compared with controls. Moreover, perhaps as a consequence, an enhanced frequency of in vivo preactivated T cells was seen as judged from an increased spontaneous proliferative response to low concentrations of exogenous IL-2. LPL-T and peripheral blood T lymphocytes exhibited similar percentages of TCR V beta gene usage both in controls and in patients. In summary, polyclonal activation of LPL-T due to impairment of local adjustment, i.e. insufficient down-regulation of TCR/CD3-dependent signalling processes, may contribute to the pathogenesis of inflammatory bowel disease.

Role of opioid peptides in the regulation of lymphocytic proliferation and in the change of the Thl/Th2-cytokinic profile

The authors studied a role of β-endorphin during the blockade of opiate receptors and selective μ- and δ-receptor agonists DAMGO and DADLE to the reaction of lymphocytic blast-cell transformation (RLBCT) and to the production of IL-1β, γ-IFN, and IL-4 in the presence of phytohemagglutinin (PHA). It was found that β-endorphin, naloxone, and the selective μ- and δ-receptor agonists stimulated PHA-induced RLBCT, without affecting a spontaneous proliferative response. During opiate receptor blockade, the effect of β-endorphin was not abolished, but, on the contrary, enhanced as naloxone exerted a stimulating effect on RLBCT. A proliferative response was not recorded during preliminary one-hour opioid treatment, β-endorphin, naloxone, and DADLW enhanced the PHA-induced production of IL-4. Naloxone exerted a heterodirectional effect on the synthesis of this cytokine, by inhibiting it spontaneously. The opioid peptides and naloxone produced no effect on the production of IL-1β) and γ-IFN. It is suggested that β-endorphin, naloxone, and the selective μ- and δ-receptor agonists promote the differentiation of Т lymphocytes towards Th2-cells.

The T-cell anergy induced by Leishmania amazonensis antigens is related with defective antigen presentation and apoptosis.

Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease associated with anergic immune responses. In this study we show that the crude antigen of Leishmania amazonensis (LaAg) but not L. braziliensis promastigotes (LbAg) contains substances that suppress mitogenic and spontaneous proliferative responses of T cells. The suppressive substances in LaAg are thermoresistant (100 degrees C/1h) and partially dependent on protease activity. T cell anergy was not due to a decreased production of growth factors as it was not reverted by addition of exogenous IL-2, IL-4, IFN-gamma or IL-12. LaAg did not inhibit anti-CD3-induced T cell activation, suggesting that anergy was due to a defect in antigen presentation. It was also not due to cell necrosis, but was accompanied by expressive DNA fragmentation in lymph node cells, indicative of apoptosis. Although pre-incubation of macrophages with LaAg prevented their capacity to present antigens, this effect was not due to apoptosis of the former. These results suggest that the T cell anergy found in diffuse leishmaniasis may be the result of parasite antigen-driven apoptosis of those cells following defective antigen presentation.

Spontaneous Peripheral T-cell Responses to the IA-2β (Phogrin) Autoantigen in Young Nonobese Diabetic Mice

Phogrin (IA-2β), a major autoantigen in type 1 diabetes in man is recognized by peripheral T cells in the nonobese diabetic (NOD) mouse. CD4+ T-cell clones derived from immunized NOD animals elicit islet destruction in a disease transfer model. Spontaneous proliferative responses to the protein and derived peptide epitopes were detected in peripheral lymph node cells (LNC) of unprimed NOD mice but not BALB/c controls as early as 4 weeks of age at a time point when insulitis in NOD animals is minimal. Responses to irradiated NOD islet cells but not irradiated NOD spleen cells were observed for both male and female NOD animals. Insulin, phogrin and phogrin-peptide 7 (aa 755–777) but not phogrin-peptide 2 (aa 640–659) or tetanus toxin peptide were recognized as antigens. Islet cell-reactive and phogrin peptide 7-specific CD4+ T-cell lines were generated from splenocytes of unprimed 4-week-old NOD females and shown to secrete Th1-type cytokines. The results show that the phogrin molecule is targeted early in the course of disease in NOD animals at a time when circulating autoantibodies are absent and insulitis is minimal.

Effects of Citrus aurantifolia concentrated extract on the spontaneous proliferation of MDA-MB-453 and RPMI-8866 tumor cell lines.

The in vitro effects of concentrated lime juice (CLJ) extract on the spontaneous proliferation of a human breast carcinoma cell line (MDA-MB-453) and a human lymphoblastoid B cell line (RPMI-8866) were investigated. CLJ extract was prepared by freeze-drying fresh fruit juice and dialyzing the concentrated extract against phosphate buffered saline in order to deplete low molecular weight micronutrients such as flavonoids as well as adjusting the pH of the extract to the physiological range. The effects of different concentrations of the CLJ extract on the spontaneous proliferative responses of the cell lines were determined by 3H-thymidine incorporation after 24 hrs of incubation. CLJ extract had no significant effect on MDA-MB-453 cell line, however, using the concentrations of 125, 250, and 500 microg/ml of CLJ extract a significant inhibition of the spontaneous proliferation of RPMI-8866 cell line was detected (P < 0.05). Due to the protein nature of the biologically active macromolecules of the CLJ extract (Gharagozloo and Ghaderi, 2001), it is reasonable to assume that the protein components of the CLJ extract may have anti-proliferative effects on tumor cell lines.

Growth Modulation of Freshly Isolated Non-Hodgkin's B-Lymphoma Cells Induced by Various Cytokines and All-Trans-Retinoic-Acid

We investigated the potential often cytokines (IL2, IL3, IL4, IL6, IL10, IL13, G-CSF, GM-CSF, interferon alpha, interferon gamma) and all-trans-retinoic acid to modulate the spontaneous proliferative response in vitro of purified B-non Hodgkin's lymphoma cells of various histological subtypes. 19 malignant lymph nodes were studied. In each case the growth could be influenced by several of these modulators. Cytokines most often implicated were interferon gamma (14/19 cases, 73.7%), IL4 (13/19 cases, 68.4%), interferon alpha (12/19 cases, 63.1%). IL2 (9/19 cases, 47.3%), IL6, IL10, IL13 and ATRA were less frequently involved (6/19 cases, 31.6%) and hematopoietic growth factors (IL3, GM-CSF, G-CSF) were rarely implicated (2/19 cases, 10.5%). The values of growth stimulation ranged from a 1.1-fold to a 6.1-fold increase, and the values of growth inhibition ranged from 15% to 98%. Each cytokine could be either inhibitory or stimulatory depending on the sample analyzed, and no relationship could be found with the histological subtype. Two notable exceptions were IL2, displaying exclusively a positive effect, and ATRA displaying exclusively a negative effect. Overall, these results may have strong implications for future clinical studies using cytokines in the treatment of lymphomas. Ideally, the pattern of in vitro growth response to cytokines or ATRA should be determined individually before undertaking any cytokine treatment.

Suppression of lymphocyte spontaneous proliferative response by proteolipid protein peptide in patients with HAM/TSP

To understand the immune mechanism suggested in HTLV-I-associated myelopathy (HAM/TSP), we investigated T cell response to proteolipid protein (PLP). Because of high autologous proliferative response (APR) of peripheral blood mononuclear cells (PBMC) in culture, the lymphocyte proliferation assay was not useful in this disease. Unexpectedly, however, APR was profoundly (70-98%) suppressed in 6 of 9 cases when PLP peptide 105-124 was added in the culture. PLP peptide 85-104 or 145-159 also suppressed APR in a few cases. Time course study showed that the peptide-mediated suppression became apparent after day 4 in culture. The results can be interpreted as that suppressor cells recognizing the PLP peptides were present in the PBMC of HAM/TSP patients and suppressed the APR as the consequence of antigen specific response. This may indicate that a T cell response to certain PLP determinants is involved in the pathomechanism of HAM/TSP at least in part. Molecular mimicry between PLP and HTLV-I may account for the T cell sensitization to PLP in HAM/TSP.

Phenotypic and functional analysis of peripheral blood lymphocytes in oral lichen planus.

To assess cellular immunity in oral lichen planus (OLP), peripheral blood mononuclear cells (PBMC) were obtained from 19 OLP patients and 30 control subjects. The proportions of circulating CD45RA+ and CD29+ lymphocyte subsets were determined. The proliferative activity of PBMC to the non-specific plant mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) was examined together with the spontaneous proliferative response and the response in the autologous mixed lymphocyte reaction (AMLR). In the OLP group, the proportion of CD4+ CD45RA+ T lymphocytes was significantly less than control subjects and the proportion of CD4+ CD29+ T lymphocytes was increased significantly. The proliferative response to PHA was similar in OLP and controls subjects. Con A-stimulated PBMC proliferation was decreased significantly in the OLP group. Spontaneous PBMC proliferation in patients with non-reticular lesions was significantly less than control subjects. Despite a mildly depressed response in the AMLR in OLP patients, this result was not statistically significant. Results of the phenotypic analysis of peripheral blood lymphocytes indicate a decreased proportion of naive T cells and an increased proportion of primed memory T cells, although the antigen specificity of these memory cells remains to be determined. Results of the functional assays would seem to reflect this phenotypic shift, and as T cells responding to Con A stimulation and in the AMLR possess suppressor-inducer activity, these results may also suggest an association between OLP and defective innate T cell-mediated suppressor circuits.

Effect of initial treatment of chronic inflammatory periodontal disease in adults on spontaneous peripheral blood lymphocyte proliferation

The spontaneous proliferative response (SPR) of peripheral blood lymphocytes, as a measure of the autologous mixed lymphocyte reaction (AMLR), has been found to be depressed in adults with chronic inflammatory periodontal disease (CIPD). The aim of the present study was to test the hypothesis that initial treatment of CIPD in adults restores the SPR to normal levels. 10 periodontal disease subjects (mean probing attachment loss of 4.2 mm and a mean bleeding index of 0.65) and 10 age- and sex-matched healthy control subjects were studied. The SPR for each patient was evaluated on days 3, 5, 7, 9 and 11 in culture, before and after initial treatment for CIPD. The peak SPR, which occurred at day 5, was depressed in the untreated periodontal disease subjects compared to the healthy control subjects (p less than 0.01). In addition, the kinetics of the SPR were found to be significantly different in 4 of the 10 parameters compared with the untreated periodontal disease patients and the healthy control subjects. After treatment, there was a significant reduction in probing attachment loss and bleeding indexes (p less than 0.001). In addition, the magnitude of the peak SPR was not significantly different from that of the healthy control subjects. Nevertheless, a difference in 1 of the 10 kinetic parameters persisted, which suggested that complete restoration of the SPR to normal had not occurred so soon after treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellular immune response to sheep erythrocytes interrelationship between proliferation of popliteal lymph node cells and footpad swelling

Mice were sensitized with graded doses of sheep erythrocytes by the intravenous or subcutaneous route and challenged for delayed-type hypersensitivity (DTH) at different times thereafter. The DTH response as assessed by footpad swelling (FPS) was compared to the spontaneous proliferative response of the popliteal lymph node cells (PLNC). Proliferation of PLNC was optimal after sensitization regimens resulting in optimal FPS. The same was true for mice sensitized under cyclophosphamide modulation. Proliferation of PLNC induced by SRBC was antigen-specific, although some crossreactivity with horse red blood cells was observed. Proliferation of PLNC could be abrogated by treatment with anti-Thy-1.2 antiserum plus complement demonstrating the T cell nature of proliferating cells. In accordance with published data, FPS of mice presensitized with a high dose of SRBC as well as FPS of recipients of spleen cells from high-dose-sensitized donors was suppressed. In marked contrast, PLNC proliferation was not diminished in these mice. Although proliferation of PLNC did not parallel FPS under all circumstances, it seems to be a correlate of the cellular immune response to SRBC.