E-rosette Formation Research Articles

Study the Ability of Enterotoxigenic E.coli Antigens To Induce Cellular Immune Response by Erythrocyte-Rosette Formation and Leukocyte Migration Inhibition Factor in Rabbits

Objectives: This study was aimed to investigate the ability of Enterotoxigenic E.coli and their enterotoxins to induce cellular immune response by means of erythrocyte-rosette formation and leukocyte migration inhibition factor tests. Methodology: Total of 30 stool sample was collected of patient with severe diarrhea whom attended to Al-Sadder teaching hospital in Al-Najaf city in period extended of April to June 2013. All bacterial isolates was identified according to morphological and biochemical testes and by API 20 E system, which further conformational tested for heat stable enterotoxin production by Suckling Mice Assay (SMA). three antigens (heat killed bacterial cell, crude enterotoxin and purified heat stable enterotoxin ETECST) was prepared and injected in experimental animals(rabbit) and tested in the level of systemic and mucosa (appendix and duodenum). Statistical analysis was done by least significant difference (LSD). Results: The results showed that 20% (6 isolates) was Enterotoxigenic E.coli and the immune experiments showed significant increase level of E-rosette formation in rabbit injected with heat killed antigen (50.1%, 49.7% and 53.0%) compared with control (23.1%, 31.8% and 29.6%) in blood appendix and duodenum consecutively, while high level appeared in crude enterotoxin and ETECST in appendix (41.3%) and (47.4%) consecutively. The assessment of ETECST LIF in migration percent to sensitizer was showing high significant LIF (54.5%, 47.6% and 43.3%) compared with control (82.0%, 79.5% and 79.5%) in blood appendix and duodenum consecutively. Conclusions: crud and partially purified enterotoxins have ability to induce cellular immune response in blood and mucosa. Recommendation: prepare a carrier molecule model for partially purified enterotoxins to make a vaccine against Enterotoxigenic E.coli .

Comparison of immunotoxic effects induced by the extracts from methanol and gasoline engine exhausts in vitro

Gasoline engine exhaust has been considered as a major source of air pollution in China. Due to lower cyto- and geno-toxicity effects of methanol engine exhaust, methanol is regarded as a potential substitute for gasoline. We have previously compared cyto- and geno-toxicities of gasoline engine exhaust with that of methanol engine exhaust in A549 cells (Zhang et al., 2007).To characterize the immunotoxic effects for gasoline and methanol engine exhausts in immune cell, in this study, we further compared effects of gasoline and methanol engine exhausts on immune function in RAW264.7 cell and rabbit alveolar macrophages. Results showed that both gasoline and methanol engine exhaust could evidently inhibit RAW264.7 cell proliferation, promote RAW264.7 cell apoptosis, decrease E-rosette formation rate and inhibit anti-tumor effects of alveolar macrophages, at the same time, these effects of gasoline engine exhaust were far stronger than those of methanol engine exhaust. In addition, gasoline engine exhaust could significantly inhibit activities of ADCC of alveolar macrophages, but methanol engine exhaust could not. These results suggested that both gasoline and methanol engine exhausts might be immunotoxic atmospheric pollutants, but some effects of gasoline engine exhaust on immunotoxicities may be far stronger than that of methanol engine exhaust.

The study on the mechanism of the inhibitory effect of the serum IgG from sarcoidosis patients on the E-rosette formation of normal T-cells (author's transl)

The study on the mechanism of the inhibitory effect of the serum IgG from sarcoidosis patients on the E-rosette formation of normal T-cells (author's transl)

Lymphocyte subpopulations in the cerebrospinal fluid and peripheral blood in multiple sclerosis.

Subpopulations of lymphocytes in the CSF and peripheral blood were studied in 30 patients with MS, 16 with other neurological diseases (OND) and 15 control subjects without any neurological abnormalities. In patients with relapse of MS, the absolute numbers of total lymphocytes, alpha-naphthyl acid esterase (ANAE) positive, E-rosette forming and bearing the "avid" FcIgG receptor lymphocytes were significantly increased in the CSF as compared with stable or slowly progressive MS patients, patients with other OND and control subjects. The relative number of ANAE-positive cells was higher, and "avid"FcIgG receptor bearing cells lower in the CSF of all patients with MS than in the two other groups. The significance of the finding is unclear. The imbalance between lymphocyte subpopulations may reflect a primary defect in MS, or may be secondary, due to the presence of circulating immune complexes. In peripheral blood no substantial differences in lymphocyte behavior were observed between MS patients and other groups.

Levamisole therapy for multiple warts.

Levamisole, a wide-spectrum antiparasitic drug, increases cellular immunity in vivo and in vitro. Patients with verruca vulgaris may have defective cell mediated immune mechanisms. Levamisole was given to twenty-two patients with multiple warts, 5 mg/kg body weight on 3 consecutive days every fortnight. Patients' lymphocytes were studied for E-rosette formation before therapy. E-rosette counts were lower in patients compared with controls and a significant increase was obtained after in vitro incubation with levamisole. Seventeen patients were cured in 1-4 months, there were four failures and one patient showed marked improvement. It is concluded that patients with multiple warts have defective cell-mediated mechanisms as far as the E-rosette count is concerned and that levamisole increases in vitro E-rosette formation and is useful in the treatment of these patients.

Documentation of immune profile of microglia through cell surface marker study in glioma model primed by a novel cell surface glycopeptide T11TS/SLFA-3.

The sheep erythrocyte membrane glycoprotein T11TS/SLFA-3 can form a ligand-receptor complex with CD2 present on immunocyte and exert stimuli for activation and proliferation. Regression of brain tumor with the application of T11TS indicates the probable role of microglia, the chief immunomodulatory cell within the brain compartment. In the present study microglial activation and immunophenotypic modulation were assessed in T11TS treated brain tumor-bearing animal models. Rat glioma models induced by chemical carcinogen ENU were treated with three consecutive doses of T11TS. Microglial cells from brain were isolated and assessed through E-rosette formation, SEM and FACS for CD2, MHC class II, CD25, and CD4. The preliminary indication of presence of CD2 on microglia through E-rosette formation was confirmed by SEM and FACS. MHC class II and CD2 single and double positive subpopulations exist, and their expression is also modulated in different doses of T11TS. A general trend of highest receptor saturation and microglial activation, measured through the activation marker CD25 and CD4 expression, was observed in 2nd dose of T11TS administration, which was then dampened via a complex immune feedback mechanism in the 3rd dose.

Influence of ethanol on CD2 receptors of lymphocytes in alcohol dependent inpatients and healthy individuals

The objective of this research was to assess the alcohol effect on E-rosette formation in vitro. The results reveal that the intoxicating dose of alcohol (0.2?-5.0?) in healthy volunteers and alcoholics' blood can differently affect the E-rosette formation: in the healthy volunteers it suppressed while in the alcoholics it stimulated the E-rosette formation. Both the suppression and stimulation were closely dependent on the alcohol consumption. Though the blood of alcoholics contained fewer E-rosette forming cells than that of the healthy volunteers, the infusion of 2.0? alcohol in vitro to the alcoholics' blood raised their E-rosette formation to the level of healthy volunteers.

Analysis of the effect of combined desensitizing acupoints therapy in treating 419 cases of allergic rhinitis accompanied asthma

Objective: To explore the effect of combined acupoint desensitizing therapy.Methods: A total of 419 cases of allergic rhinitis accompanied with asthma were treated by combined acupoint desensitizing therapy with positive allergen extract on acupoints of head and upper back region.Results: After 3 courses of treatment, skin test repeated with responding allergen extract showed that the scope of redness and swelling of skin were smaller than those before treatment,P < 0.01. The3H-TdR incorporation lymphocyte transformation rate, acidophil count, IgA, IgG level and E-rosette formation rate of acupoint desensitizing group were different significantly from those of the two control groups (the acupoint 0.85% normal saline injection group and the subcutaneous desensitizing group),P < 0.01 orP < 0.05. Follow-up study for over 3 years showed that the markedly effective rate of the acupoint desensitizing group was 68.73%, the effective rate 29.12% and total effective rate, 97.85%. The effect was higher than that of the two control groups, P<0.01.Conclusion: Combined acupoint desensitizing therapy is valuable and worthy of popularizing in treating patients of allergic rhinitis accompanied asthma.

Influence of ethanol on E-rosette formation by T lymphocytes in alcoholics: Preliminary report

The immunopathology of patients with alcohol dependence has been shown earlier. The purpose of the present study was therefore to investigate the influence of alcohol on the E-rosette-forming reaction of lymphocytes in alcoholics, to ascertain whether alcohol dependence is indicated by the functioning of the human immune system. Nineteen patients with alcohol dependence and 11 healthy persons were investigated. Ethanol at a concentration of 0.02% and more was found to inhibit the formation of E-rosettes by lymphocytes in healthy persons and to stimulate their formation by lymphocytes in alcoholic patients.

Effect of ribonuclease from Bacillus intermedius on human blood lymphocytes

By using a rosette formation test the effect of ribonuclease Bacillus intermedius (RNase Bi) on T- and B-lymphocytes in human peripheral blood has been studied in vitro. The RNase effect on T-lymphocytes depends on its concentration: low concentrations (10 −6–10 −2 μg ml −1) stimulate E-rosette formation whereas high concentrations (10 μg ml −1) suppress it. The amount of B-lymphocytes decreases under RNase Bi influence in all concentrations tested. RNase Bi like thymus hormones influence immature lymphocytes (0-cells) by inducing the surface expression of E-receptors what leads to rosette formation and, thus, contributes to lymphocyte differentiation. The increase in the amount of active T-cells which represent the mature cell population also confirms the participation of RNase Bi in T-rank lymphocytes differentiation processes. The RNase Bi effect on the human lymphocytes depends on its catalytic activity.

Effect of ribonuclease from Bacillus intermedius on human blood lymphocytes.

By using a rosette formation test the effect of ribonuclease Bacillus intermedius (RNase Bi) on T- and B-lymphocytes in human peripheral blood has been studied in vitro. The RNase effect on T-lymphocytes depends on its concentration: low concentrations (10(-6)-10(-2) microg ml(-1)) stimulate E-rosette formation whereas high concentrations (10 microg ml(-1)) suppress it. The amount of B-lymphocytes decreases under RNase Bi influence in all concentrations tested. RNase Bi like thymus hormones influence immature lymphocytes (0-cells) by inducing the surface expression of E-receptors what leads to rosette formation and, thus, contributes to lymphocyte differentiation. The increase in the amount of active T-cells which represent the mature cell population also confirms the participation of RNase Bi in T-rank lymphocytes differentiation processes. The RNase Bi effect on the human lymphocytes depends on its catalytic activity.

Haemocytes in Cerastoderma edule (Mollusca, Bivalvia): distinct cell types engage in different responses to sheep erythrocytes

Three types of circulating haemocytes were identified in the haemolymph of the cockle Cerastoderma edule with light microscopy (LM), and electron microscopy (EM) and enzyme cytochemistry. Type I cells are large spreading haemocytes, with a ruffled surface, eccentric nucleus and distinct cytoplasmic organelles. These cells are esterase- and acid phosphatase- positive. Type II cells are spherical, with a smooth surface and central nucleus surrounded by a cytoplasm devoid of organelles. They are negative for esterase and acid phosphatase reactions. Type III cells, which have not been described before in molluscs, have a round shape, a small eccentric nucleus and a large vacuole filled with granular material (TEM) occupying the majority of the cytoplasm. These cells are peroxidase positive and morphologically resemble the serous cells of bivalves or the signet ring cells of urochordates. The cellular response of Cerastoderma edule to the intracoelomic injection of sheep red blood cells (SRBC) was studied. The cell types involved in E-rosette formation and phagocytosis were documented by LM, TEM and SEM. In light microscopy and ultrastructural analysis the E-rosette forming cell was identified as Type II cells. The Type I cells phagocytosed SRBC, autologous cells and remnants thereof. Red blood cells with disrupted membranes and different electron densities were observed not only inside but also in close vicinity of phagocytic cells, which may suggest that the cells produce a cytolytic factor. The function of Type III cells remains obscure.

Persistent Reduction of Complement Receptor 3 α-Chain Expressing Mononuclear Blood Cells and Transient Inhibitory Serum Factors in Whipple's Disease

Several small studies have indicated an impaired cell mediated immune response as a possible cause for the delayed elimination of the bacteria in Whipple's disease. A specific defect, however, has not been defined. We examined the expression of cell surface molecules and mitogenic responses of peripheral blood mononuclear cells in 27 patients with Whipple's disease at different disease stages by indirect immunofluorescence and by measurement of [ 3H]thymidine incorporation, respectively. E-rosette formation and cutaneous reaction to seven recall antigens were determined. Matched healthy donors served as controls. We found a significantly reduced number of cells expressing the complement receptor 3 αchain (=CD11b) in all patients. In florid disease, the number of activated cells (in particular CD58 positive cells) was increased and CD4/CD8 ratios were diminished. Proliferation to phytohemagglutinin and to sheep red blood cells was reduced at all stages of the disease. Serum of control persons reversed this decreased responsiveness especially in patients with active disease. Skin reaction was hypoergic in all patients. Determination of CD58 positive cells increased in patients with active disease may be useful to define the activity of the disease and the duration necessary for treatment. Transient inhibiting serum activities may impair the CD2/CD58 interaction. The reduction of cells expressing CD11b, the decreased proliferation, and the cutaneous hypoergy indicate a persisting defect of cell mediated immunity in vivo and in vitro. These defects may contribute to the impaired ability of patients with Whipple's disease to eliminate bacteria.

Myelosuppression in HCL: Role of hairy cells, T cells and haematopoietic growth factors

To elucidate mechanisms which may be responsible for the haematopoietic insufficiency in hairy cell leukaemia (HCL), we investigated in an autologous in vitro system the influence of haematopoietic growth factors (CSFs) and the effects of hairy cells (HCs) as well as T cells on the formation of haematopoietic colonies (CFU). Colony forming assays were performed using peripheral blood mononuclear cells (PBMC) of 6 HCL patients. To remove HCs, PBMCs were subjected to complement-mediated lysis, T cells were removed by E-rosette formation. Assays were done with and without recombinant human (rh) interleukin-3 (IL-3) and rh granulocyte-macrophage-colony-stimulating factor (GM-CSF). All 6 patients exhibited a severe reduction of their circulating progenitor cell (CPC) compartment. There was no correlation between the degree of colony reduction and the number of HCs. However, a correlation was found between the numbers of CPCs of HCL patients and healthy donors and the monocyte counts in these groups (r = 0.8573, p < 0.001). The removal of autologous HCs, but also of T cells, resulted in a significant increase in colony formation (BFU-E, CFU-GM, CFU-mix). In none of the experiments, however, did colony numbers come close to the normal range. This was only achieved by supplementation of the culture medium with rh IL-3 and rh GM-CSF. The results suggest that the haematopoietic failure observed in HCL patients is probably due to an inadequate supply of CSFs as well as to an inhibitory activity of HCs and T cells which might exert their effects in a synergistic fashion. There is also evidence that the lack of monocytes plays a role in the development of the haematopoietic insufficiency in HCL.

Characterization of a new porcine differentiation antigen involved in the proliferative response to mitogens

The cytotoxic monoclonal antibody (Mab) LIL 13 reacts with a widely distributed antigen that is expressed on 80–95% of swine peripheral blood mononuclear cells (PBMC) and a variety of lymphoid cell types. Using indirect immunofluorescence (IIF) the positive cells (75–100%) were divided between the bright, intermediate and dull populations. The remaining negative cell population contained B-cells, T-cells and probably null cells. Mab LIL 13 did not react with swine major histocompatibility complex (MHC) class I antigens (SLA) and did not inhibit E-rosette formation. Reactivity of LIL 13 with leukocyte function antigen 1 (LFA-1) was excluded by competitive IIF and cytotoxicity tests with cross-reacting anti-human CD 18 or anti-swine LFA-1-specific antibodies. Mab LIL 13 and complement treatment severely reduced mitogen-induced proliferative response to phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) and lipopolysaccharide (LPS) (90–100%). In the absence of complement, LIL 13 partially reduced proliferation of cells by interfering with the capability of mitogens to bind to the corresponding surface receptor (LIL 13 followed by mitogens), and partially inhibited mitogenic proliferative response following post-treatment (mitogens followed by LIL 13). Biochemical analysis of the antigen using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed bands of approximately 180–190 kDa and 46–50 kDa under reducing conditions and 200 kDa and 46–50 kDa under non-reducing conditions.

DEVELOPMENT AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO LYMPHOCYTE CELL SURFACE ANTIGENS OF THE RHESUS MONKEY

Three monoclonal antibodies directed against rhesus lymphocyte cell surface antigens are described. A pan-T mAb, T64, and a T suppressor mAb, T35, showed phenotypic and functional specificity for both human and rhesus cells. In contrast, a third mAb, N42, identifying natural killer cells in rhesus peripheral blood leukocytes, was not crossreactive with the corresponding homologous human cells. N42 reacted with the same cells identified by Leu 11a and Leu 11b in rhesus PBL and in a functional assay decreased NK activity by 80%. N42 precipitated a 50KD protein from rhesus PBL lysates and was reactive with the Fc receptor domain of the NK cell. The T-S functional activity of cells reactive with mAb T35 was demonstrated in a pokeweed-mitogen-driven system for Ig synthesis: removal of the T35 positive cells by complement-mediated lysis led to an enhanced production of Ig by rhesus PBL, and the addition of T35 positive cells to a culture of T helper and B cells resulted in a reduction of this response. T35 was determined to be an IgG2a immunoglobulin and precipitated a 34KD protein from rhesus cell lysates. An IgM immunoglobulin, mAb T64 delineated all T lymphocytes, inhibited E-rosette formation, interfered with the proliferation of cells stimulated with mitogens, and precipitated a 52KD membrane protein. The potential utilization of these mAbs in vivo for organ or tissue transplantation in the rhesus monkey is discussed.

Participation of cytoplasmic organelles in E‐rosette formation

To elucidate the mechanism of E-rosette formation between T cells and sheep red blood cells (SRBC), the effect of various agents affecting the function of cytoplasmic structures (microtubules and microfilaments) and organelles (mitochondria and rough endoplasmic reticulum) was investigated. E-rosette formation was greatly inhibited by agents that block either the energy producing system (KCN and NaN3) or the integration of microfilaments (cytochalasin B, dihydrocytochalasin B and cytochalasin D). On the other hand, there was little or no suppression by either inhibitors of protein synthesis (cycloheximide and puromycin), agents that block the polymerization of microtubules (colchicine, podophyllotoxin and vinblastine), or chemicals disconnecting surface membrane proteins from the intracellular structural proteins (chlorpromazine, dibucaine, hydrocortisone, and propranolol). The calcium ionophore A23187, which transports Ca2+ into the cytosol, inhibited the E-rosette formation in the presence of Ca2+, but not in its absence. From these results, we concluded that new synthesis of ATP and the structural integration of microfilaments are indispensable for the E-rosette formation, which is triggered by an interaction between the ligand (T11TS) and its corresponding receptor (CD2). A certain level of intracellular Ca2+ is also involved in the E-rosette formation.

Studies on T‐Lineage Cells in Human Decidua of First Trimester Pregnancies

T-lineage cells in human decidua of early pregnancies were tested for surface markers, proliferative response, interleukin-2 (IL-2) production, and natural killer (NK) activity. T-lineage (CD2+) cells that were obtained from decidua by the use of E-rosette formation contained fewer CD3+ mature T cells and CD4+ cells than those from the peripheral blood of the same donors, while no differences were seen in the frequencies of CD8+ cells. P55 molecules of IL-2 receptor (IL-2R/p55, Tac antigen) were hardly detected on fresh decidual T-lineage cells, though approximately 20% were positive for HLA-DR. More than a half of decidual T-lineage cells expressed CD56 molecules on their surface and killed K562 cells, the prototype target of NK cells, while most of them were negative for CD16 and CD57. Upon stimulation with IL-2, decidual T-lineage cells demonstrated dose-dependent proliferative response. In addition, they were induced to produce high amounts of IL-2 by stimulation with mitogens but not with alloantigens. These results suggest that human decidua contains high numbers of CD2+3-CD16 +/- 56+ lymphocytes and that this population responds to IL-2, produces IL-2 and mediates NK activity.

In vitro studies of Tyr-MIF-1 with human lymphocytes

Our previous report showed that the brain peptide Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH 2) blocks the inhibitory effect of morphine sulfate on E-rosette formation by human peripheral blood lymphocytes (PBL). In this study, additional in vitro effects of Tyr-MIF-1 on human PBL were studied. The percentages of positive cells for CD 2, a sheep erythrocyte receptor, CD 4 and CD 8 were unchanged after incubation of PBL with morphine or morphine plus Tyr-MIF-1. Tyr-MIF-1 was not mitogenic by itself. The addition of Tyr-MIF-1 did not increase the proliferative response of PBL to Con A, although morphine did. Tyr-MIF-1 did not activate PBL to produce IL 2 nor did it affect the production of IL 2 by Con A-stimulated PBL. The results suggest that Tyr-MIF-1 does not directly modulate CD 2, CD 4 and CD 8 expression, does not alter the proliferative response of PBL, and does not affect the production of IL 2.

Some immunological findings in adult periodontitis.

There is little doubt that immunological mechanisms play an important role in chronic inflammatory periodontal disease. At the same time, it is recognized that patient susceptibility is ultimately responsible for the clinical manifestation of the disease. In this context, the present study was undertaken to examine a range of systemic immunological parameters in patients with adult periodontitis (AP), so as to test the hypothesis that a specific pattern would identify diseased--possibly 'at risk'--patients. These parameters included serum IgA, IgG, IgM, IgD, C3, transferrin, the presence of circulating immune complexes, and the number of circulating T (E-rosette forming) cells. One hundred and forty AP patients and 70 healthy controls were examined. Following a complex statistical analysis only the levels of IgG, IgM and IgD were significantly increased in adult periodontitis (p less than 0.05) while an increase in circulatory immune complexes was significant only for separate statistical tests. Although statistically different, the levels seen in AP patients were still within the normal range hence the clinical significance of the findings is such that it is unlikely that these systemic immunological parameters per se do define an 'at risk' population.