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Seasonal effects of rotational stress on Lewis lung carcinoma metastasis and T-lymphocyte subsets in mice.

Rotational stress specifically increases the formation of spontaneous lung metastasis in mice bearing Lewis lung carcinoma, without significantly modifying the growth of primary tumor. The increase in metastasis number and volume caused by rotational stress varies in magnitude with a highly significant circannual rhythm; the acrophase approximately coincides with summer solstice. Rotational stress causes a significant reduction in the number of CD3+ and CD4+ T-lymphocyte subsets in summer, whereas in winter the number of CD3+ subset is significantly increased; the CD4+/CD8+ ratio and the number of NK 1.1 antigen positive cells are not significantly modified by rotational stress in both periods considered. The increase in metastasis formation by rotational stress thus appears to negatively correlate with the number of splenic CD3+ and CD4+ T-lymphocyte subsets. This seasonal behavior occurs in spite of the control of light cycle, temperature and humidity in the animal housing, suggesting the existence in the host of an endogenous oscillator with a circannual period. These data indicate the opportunity to consider endogenous rhythms within the host, as well as seasonal factors, in studies on stress and neuroimmunomodulation in experimental oncology.

Analysis of Variance↗

[Reference values of T-lymphocyte subsets in umbilical cord blood].

In order to determine reference values of T lymphocyte subpopulations in cord blood in the metropolitan area of Puebla city we randomly selected 80 from a total of 400 term, eutrofic and without pathology newborn. The percentage of CD3, CD4 and CD8 T lymphocyte subpopulations in cord blood were determined by flow cytometry. Reference values were gotten from the 25 and 75 percentile in parameters without gaussian distribution. Reference values are CD3 (45.5 to 91.3%), CD4 (26.2 to 69.4%), CD8 (16.2 to 24.1%) and NK (2.0 to 8.5%). The presented data represent valuable normal ranges of T lymphocyte subsets to compare with values observed in sick or in risk of developing immunoallergic pathology children.

Cross-Sectional Studies↗

Unique order of the lymphocyte subset induction in the liver and intestine of mice during Listeria monocytogenes infection.

We investigated how NK cells, extrathymic T cells, and thymus-derived T cells are activated in mice during infection with an intracellular pathogen, Listeria (L.) monocytogenes. Although macrophages and granulocytes are known to be involved in the elimination of this pathogen in an early phase of infection, it was still controversial what type of lymphocytes are induced as effectors in subsequent phases. When mice were ip injected with 1 x 10(3) L. monocytogenes (a sublethal dose), a prominent increase in the number of mononuclear cells in the liver and spleen was induced. Phenotypic analysis revealed that serial induction of lymphocyte subsets, NK cells-->extrathymic T cells-->thymus-derived T cells, occurred in these organs. Extrathymic T cells were estimated to have intermediate CD3 and a high level of IL-2 receptor beta-chain on the surface (i.e., intermediate CD3 cells). These mice became free from infection after 2 weeks. In the case of oral administration, 1 x 10(3) L. monocytogenes increased the number of cells in the liver and the number of intraepithelial and lamina propria lymphocytes in the intestine. Phenotypic analysis also showed a sequential induction of lymphocyte subsets in the liver and the induction of extrathymic T cells in the intestine. Preelimination of intermediate TCR cells and NK cells by in vivo treatment with anti-LFA-1 mAb made mice susceptible to an ip injected sublethal dose of L. monocytogenes. These results reveal a unique order of lymphocyte induction during listerial infection and indicate that extrathymic T cells might be one of the important cells in achieving resistance against L. monocytogenes.

Animals↗

Fixation and cryopreservation of whole blood and isolated mononuclear cells: Influence of different procedures on lymphocyte subset analysis by flow cytometry.

BACKGROUND: Immunophenotyping of whole blood (WB) and isolated peripheral blood mononuclear cells (PBMCs) is a common tool used to evaluate immune system changes in clinical studies. The development of methods that would allow preservation of samples for flow cytometric analysis is important for the extension of this technology to field testing in settings where the equipment might be not readily accessible. METHODS: Three-color flow cytometric analysis was used to determine percentages of T cells and their subsets (CD3(+), CD4(+), CD8(+)), B cells (CD19(+)), and natural killer cells (CD16(+)/56(+)) in WB and PBMCs using variations of a standard stain/fix WB staining procedure (Optilyse) that included staining following fixation and freezing of fixed samples before or after staining. RESULTS: Comparable lymphocyte subset percentages in WB or PBMCs were observed regardless of Optliyse method used (all Ps >/= 0.8). However, differences in fluorescence intensity for several markers were observed across procedures. Compared with the standard stain/fix procedures, fix/stain decreased the mean fluorescence intensities for CD4, CD8, CD19 and CD16/56 in WB and PBMCs (P </= 0.03 for these markers P = 0.105 for CD8 in PBMCs). Further decreases in mean fluorescence intensity were seen with the fix/stain/freeze procedure. The stain/fix/freeze yielded intensities largely comparable to those seen with standard stain/fix procedure (P >/= 0.13), suggesting that, when the markers of interest are known at the time of field collection, implementation of this procedure might be desirable. Fix/freeze/stain resulted in diminution of intensity in general, but they tended to be more modest than those seen for fix/stain/freeze and therefore might be applicable to field studies in instances when the specific markers of interest cannot be defined upfront. CONCLUSIONS: Freezing of fixed WB and PBMCs before or after cell surface staining is a reliable method for preserving specimens in field sites for later determination of lymphocyte subset percentages, which are commonly assessed in immunodeficient and cancer patients.

Blood Preservation↗

Study of serial lymphocyte subsets in multiple sclerosis: their possible role in the evaluation of disease progression.

We followed-up 30 patients with definite multiple sclerosis (MS) for one year recording monthly: clinical evolution, neurological examination and T-lymphocyte subsets in peripheral blood recognized by monoclonal antibodies. Although clinical relapses could not be strictly associated with lymphocyte abnormalities in all cases, the parallel clinical-immunological survey permitted to correlate significantly the type of clinical course (chronic-progressive, relapsing, stable) with a correspondent immunological pattern (steadily high, transiently high, steadily normal CD4/CD8 ratios respectively). Our results suggest a possible role of serial lymphocyte subsets analysis in defining evolution of MS.

Adult↗

Lymphocyte proliferation assay in response to Borrelia burgdorferi in patients with Lyme arthritis: analysis of lymphocyte subsets.

The objective of this study was to assess the validity of the lymphocyte proliferation assay in the diagnosis of Lyme arthritis (LA). We analyzed peripheral blood mononuclear cells of 11 patients with LA and 5 healthy controls for proliferative responses to live Borrelia burgdorferi by 3H-thymidine uptake. Before and after proliferation, the total number of cells was estimated and the relative amount of lymphocyte subsets was determined by FACS Lymphoproliferative responses to B. burgdorferi in patients with LA were predominantly due to CD4+ cells and in a small part due to CD8+ cells and were antigen-dependent in comparison to healthy donors. Considering the number of cells before proliferation, responses of TCR gamma/delta + cells and CD56+ cells were higher than responses to TCR alpha/beta+ and CD3+ cells in LA patients, but not in healthy donors. There were no demonstrable antigen-dependent CD19+ cell responses. Our results support the validity of lymphocyte proliferation assay in the diagnosis of LA.

Adolescent↗

Comparison of lymphocyte subsets, monocytes, and NK cells in three different lung compartments and peripheral blood in the rat.

Investigations on leukocyte populations in the lung have shown that lymphocytes are found in different anatomical compartments. Lymphocytes can be seen to a different extent in the lung interstitium, the epithelium and lamina propria of the bronchi, the bronchoalveolar space, and the marginal lung vascular bed. Previous studies focused on one compartment only, or a mixture of leukocytes from lung homogenates were prepared. This study compared cellular yields from the lung parenchyma, the bronchoalveolar space, and the perfusate of the lung vasculature of healthy male Lewis rats. All compartments were investigated in the same animal, and seven different lymphocyte subsets, monocytes, and natural killer (NK) cells were analyzed using flow cytometry. It was found that the perfusate contained a high proportion of CD4+ lymphocytes compared to the lung interstitium. A very high proportion of CD4+ lymphocytes in the bronchoalveolar lavage (BAL) expressed markers for "memory" T cells. Compared to the blood, the percentage of B and T cells was much lower in the perfusate, whereas the NK cells and monocytes were more frequent. Analysis of leukocyte subsets within all compartments revealed specific, distinguishable cell compositions. Extraction of interstitial lung cells was performed using two different methods. Enzymatic digestion of the lung tissue was compared with a mechanical disruption method. Hardly any differences were observed between the two methods regarding the distribution of lymphocyte subsets, monocytes, and NK cells. These data document the need to study more than one compartment before extrapolating to lymphocytes in the lung in general. Furthermore, changes in numbers of leukocytes and subsets can now be studied in models of lung infections and immune reactions, including the entry from the blood and intrapulmonary migration from one lung compartment to the other.

Animals↗

Intrathyroidal and circulating lymphocyte subsets in different stages of autoimmune postpartum thyroiditis.

Postpartum thyroiditis (PPT) is a reversible form of lymphocytic thyroiditis which has been attributed to an aggravation of preexisting subclinical autoimmune thyroiditis. In this study no differences in circulating lymphocyte subsets were found between 9 thyrotoxic and 18 hypothyroid PPT patients and normal subjects. We obtained sufficient numbers of thyroid-infiltrating lymphocytes for surface marker characterization in 3 women in the thyrotoxic phase and in 10 women in the hypothyroid phase of PPT. Cells were identified by conventional T and B cell markers as well as by monoclonal antibodies (OKT) directed against different T cell subsets in a microscale immunofluorescence assay. In the hypothyroid patients a relative accumulation of B cells (31% vs. 17%; P less than 0.01 by the Wilcoxon signed rank test) was found within the thyroid when compared to peripheral blood. A relative decrease in intrathyroidal supressor-cytotoxic (OKT 8+) T cells (19% vs. 28%; P less than 0.01) resulted in an increased intrathyroidal helper to suppressor-cytotoxic (OKT 4+/OKT 8+) T cell ratio (3.0 vs. 2.0; P less than 0.01). Intrathyroidal lymphocyte subsets in the thyrotoxic patients were comparable to those in the hypothyroid patients. These findings, which are similar to those we previously obtained in patients with chronic Hashimoto's thyroiditis, may indicate that local synthesis of thyroid-directed autoantibodies is of primary importance in all stages of autoimmune thyroiditis.

Adult↗

Alterations in lymphocyte subsets and pituitary-adrenal gland-related hormones during fasting.

We investigated changes in the immunoendocrine system during fasting. Ten hospitalized patients aged 14-46 y with psychosomatic disorders fasted for 7 or 10 d. Blood samples were collected before and on days 3 and 7 of the 7-d fasts. When fasting continued to 10 d, an additional sample was taken on day 10. We measured blood cellularity (white blood cells and total lymphocytes), the total number and percentage of lymphocyte subsets (CD2, CD3, CD4, CD8, and CD19), natural killer (NK) cell activity, cytokines (interleukin 1 beta, interleukin 2, interleukin 6, granulocyte-macrophage colony stimulating factor, tumor necrosis factor alpha, and interferon gamma), and soluble interleukin 2 receptors. Corticotropin, cortisol, and dehydroepiandrosterone sulfate (DHEAS) concentrations were also determined. Although the total number of lymphocytes decreased during fasting, NK cell activity increased significantly. Plasma cortisol and DHEAS concentrations also increased significantly whereas changes in corticotropin concentrations were not significant. The total number and percentage of CD4 cells decreased significantly during fasting but no other lymphocyte subsets changed significantly. The percentage of CD4 cells was negatively correlated with cortisol concentrations during fasting. No detectable changes occurred in cytokines or soluble interleukin 2 receptors during the study. All measured immunoendocrine values that changed during fasting returned to prefasting values during the refeeding period. These findings indicate that fasting affects immune variables such as T cell subsets and NK cell activity at least in part through changes in adrenal gland-related hormones.

Adolescent↗

Lymphocyte subsets and activation markers in patients with acute episodes of idiopathic anaphylaxis.

BACKGROUND: Idiopathic anaphylaxis (IA), a type of anaphylaxis in which no external allergen can be identified, is a corticosteroid-responsive disease, that suggests that it may have an immunologic pathogenesis. OBJECTIVE: The objective of this study is to compare patients with acute episodes of IA with normals, patients with chronic idiopathic urticaria, and patients with IA in remission relative to lymphocyte subsets and activation markers. METHODS: This is a prospective cohort study of 38 adults: 5 normals, 4 idiopathic urticaria, 11 IA patients in remission, 9 IA patients with acute attacks who had not yet received prednisone, and 9 IA patients who had received prednisone. The main outcome measures were lymphocyte subset and activation markers determined by two and three color flow cytometry (CD2, CD3, CD4, CD5, CD8, CD16, CD19, CD23, CD25, CD56, and HLA-DR). RESULTS: Comparing patients with acute IA with those in remission, the only significant difference was that the acute IA patients had a significantly higher percentage of CD3+HLA-DR+ cells. Normals had a significantly lower percentage of CD3+ HLA-DR+ cells than all other groups. Patients with acute IA on prednisone as well as IA patients in remission had a significantly higher percentage of CD 19+ CD23+ cells than normals. CONCLUSIONS: These results suggest that there are more activated T cells in patients with acute episodes of IA than in patients in remission. Perhaps, these activated T cells have a role in the pathogenesis of IA.

Anaphylaxis↗

Modification of lymphocyte subsets in patients with rhinoscleroma.

PURPOSE: Rhinoscleroma is a rare, chronic, granulomatous disorder of the upper airways. This disease presents some etiopathogenetic aspects that are not yet clear. Infection by Klebsiella rhinoscleromatis is fundamental for the onset of the disease, but it is impossible to reproduce rhinoscleroma experimentally only via infection with the bacteria both in man and in animals. Furthermore, this disease mainly affects blood-related people and occurs in certain geographic areas. In this context, we present a study that brings to light some of the quantitative abnormalities of the lymphocyte subsets. MATERIALS AND METHODS: The study group consisted of 5 patients with rhinoscleroma. The following parameters were studied for each patient: clinical manifestations, histologic examinations, number of leukocytes, lymphocytes, and lymphocyte subsets. RESULTS: In all patients, we noted the following: There was a relative reduction of the CD4+ cells, an absolute increase of the CD8+ cells, and an inversion of the CD4+/CD8+ ratio. There was an absolute increase of the CD56+ cells and cytotoxic cells that coexpress CD8+CD56+ antigens. There was a relative reduction of the CD3+ cells, and the CD19+ cells tended to show an ambiguous behavioral pattern. CONCLUSION: We believe that K. rhinoscleromatis does not play a major role in the etiopathogenesis of rhinoscleroma. However, we do believe that the anomalous behavior of the immune system can favor rhinoscleroma.

Adult↗

Effects of low frequency electromagnetic fields on expression of lymphocyte subsets and production of cytokines of men and women employed in a museum.

The objective of this study was to analyse the immune response to electromagnetic fields (ELMFs) in seven men and eight women employed in a museum. The workers were exposed in a room to an ELMFs (range 0.2-3.6 microT and 40-120 V/m) induced by 50 Hz electricity for 20 h a week. Control groups consisted of 47 women and 39 men with a similar percentage of atopic subjects, age (range 30-51 years) and smoking habits of the workers included in the study. Levels of blood lead (Pb) and urinary trans-trans muconic acid, a metabolite of benzene (markers of exposure to traffic and smoking) of the control and exposed groups were similar. Lymphocyte subsets were determined in men and women using conjugated antibodies. Serum interleukin (IL) 4 and interferon gamma and their 'in vitro' production by peripheral mononuclear blood cells (PMBCs) stimulated by phytohemoglutinin (PHA), as well as blastogenesis of PMBCs induced by PHA, were determined in women only. ELMF-exposed women showed a significant reduction in the percentage of B and NK CD3(-)-CD25+ lymphocytes and a slight reduction of CD16(+)-56+ NK lymphocytes. They also showed significantly lower levels of interferon gamma in serum, or produced in the supernatants by PMBCs both spontaneously and stimulated by PHA, while they did not show significant changes in serum and 'in vitro' produced IL-4, or in blastogenesis of PMBCs. Men working in the museum showed, in relation to the controls, a statistically significant reduction in both number and percentage of CD16(+)- CD56+ and CD3(-)-CD25+ lymphocyte subsets. On the whole, this investigation demonstrates a reduction of blood NK lymphocytes and of the production of interferon gamma in workers exposed to low frequency ELMFs. Recent studies have shown that stress and poor lifestyle induce the reduction of blood cytotoxic activities possibly acting on nervous functions. This may suggest that ELMFs reduces blood NK lymphocytes by combined effects on the immune and nervous systems.

Adult↗

Effect of hormone replacement therapy on spinal bone mineral density and T lymphocyte subsets in premature ovarian failure and Turner's syndrome.

OBJECTIVE: Our purpose was to evaluate the effect of hormone replacement therapy (HRT) on bone metabolism and the immunological change in premature ovarian failure (POF) and Turner's syndrome. METHOD: The study was conducted on 17 POF patients, 10 Turner's syndrome patients and 35 control subjects aged 20-40 years. Bone mineral density (BMD) of lumbar vertebra by dual energy X-ray absorptiometry, serum bone metabolic parameters and T lymphocyte subsets in the peripheral blood were investigated. RESULT: The untreated and treated patients of POF (0.829 +/- 0.077, 0.918 +/- 0.094 g/cm2) and the untreated and treated patients of Turner's syndrome (0.647 +/- 0.037, 0.885 +/- 0.148 g/cm2) showed spinal osteopenia compared with the control group (1.039 +/- 0.107 g/cm2). They showed a significantly lower percentage of CD4 and a ratio of CD4/CD8 in T lymphocytes as compared with the control group before HRT. CONCLUSION: POF and Turner's syndrome patients showed marked spinal osteopenia and abnormal T lymphocytes subsets. HRT slightly improved their BMD and immunological status.

Absorptiometry, Photon↗

Identification of lymphocyte subsets and natural killer cells in head and neck cancers. An immunohistological study using monoclonal antibodies.

We investigated host-immune defenses against head and neck cancer cells by using various monoclonal antibodies with an immunoperoxidase technique to define lymphocyte subsets and natural killer (NK) cells. By so doing, we were able to identify lymphocyte subsets and NK cells in various head and neck cancers. We found that the majority of these cells infiltrate in or around nests of cancer cells and are stained with Leu-1 antibody. They include both Leu-2a and Leu-3a positive cells, which show equally intense levels of infiltration. Leu-7 positive cells were only scattered in the peripheral portion of the cancer nests in some cases. We also found a tendency for T-cells to infiltrate more intensely in poorly differentiated squamous cell carcinomas (SCC) than in moderately or well-differentiated SCC. Similarly, T-cells were more prevalent in maxillary carcinomas than in laryngeal carcinomas. These findings suggest the presence of a host-immune defense mechanism against cancer cells in patients with head and neck cancers.

Adult↗

Interpreting data on lymphocyte subsets in the blood of HIV patients - organ distribution, proliferation and migration kinetics are critical factors.

Lymphocytes stay in the blood only a short time before migrating to lymphoid and nonlymphoid organs. They represent only about 2% of all lymphocytes in the body. The ratio of CD4+/CD8+ lymphocytes in the blood depends on age and genetic influences. In HIV infection not only relative but also absolute numbers of lymphocyte subsets should be determined. The different effects of proliferation and apoptosis on lymphocytes in HIV infection have to be considered. Lymphocyte levels in the blood of HIV patients do not mirror alterations in the lamina propria of the gut or lymph nodes. The dynamic aspects of lymphocyte life span and migration during HIV infection and the progression to AIDS as well as the effects of treatment have to be taken into consideration to enable a meaningful interpretation of experimental data. More data from animal models of HIV infection are needed to study these kinetic aspects.

Age Factors↗

Changes in the expression of surface receptors on lymphocyte subsets in the elderly: quantitative flow cytometric analysis.

The immunophenotype of circulating lymphocytes, including the intensity expression of surface receptors, changes with ageing. Until now, no results of systematic studies on age-dependent changes with respect to the expression of the major lymphocyte surface receptors in healthy elderly subjects have been reported. In order to identify age-related changes in both representation and immunophenotype of lymphocyte populations, we investigated, by means of triple-color whole-blood immunostaining and quantitative flow cytometry, the percent values and the absolute numbers, as well as the levels of surface antigen expression or antigen molecules per cell (ABC values x 10(3)), of different peripheral blood lymphocyte subsets from 23 healthy elderly subjects and 13 young donors. Naive (CD45RA+CD3+) T cells, total B cells, and CD5+ B lymphocytes are decreased (22%, 6%, 0.8% vs. 30%, 12%, 1.4%, respectively), whereas activated (HLA-DR+CD3+) and memory (CD45RO+CD3+) T cells, CD3+CD7- T lymphocytes, and lymphocytes expressing the NK marker CD56 are expanded in the elderly (2%, 53%, 13%, 6% vs. 0.8%, 45%, 8%, 8%, respectively). Moreover, T lymphocytes from elderly individuals express lower CD3 (61 +/- 10) compared to young (69 +/- 10). Considering the different T-cell populations, CD3 antigen is respectively decreased on CD45RO+ T cells (55 +/- 14 vs. 66 +/- 14) and up-regulated on CD56+ T lymphocytes (62 +/- 21 vs. 45 +/- 20). Increased CD8 expression characterizes CD3+CD7- lymphocytes (70 +/- 34 vs. 44 +/- 17) while HLA-DR on activated T cells is lower in old (39 +/- 7) than young (46 +/- 9) donors. CD7 is down-regulated both in T (22 +/- 3 vs. 28 +/- 3) and NK (48 +/- 18 vs. 71 +/- 18) cells, whereas CD2 expression, unchanged on NK cells, is up-regulated on T lymphocytes (54 +/- 10 vs. 41 +/- 8). Age-related changes in B-cell antigen expressions were also found: CD20 is increased (124 +/- 23 vs. 105 +/- 16) whereas, despite the unchanged CD5 expression of T cells, CD5 intensity on the B-cell subset co-expressing this antigen is higher in old (49 +/- 37) than in young (22 +/- 4) people. The observed changes in the expression of functionally important cellular receptors can contribute to the remodeling of immune function characteristic of the elderly. Moreover, since quantitative flow cytometry is becoming widely employed in clinical practice, our results also contribute to the assessment of specific age-dependent antigen expression changes to be considered for diagnostic approaches in the elderly.

Adult↗

Immunoglobulin levels and lymphocyte subsets following cardiac operations: further evidence for a T-helper cell shifting.

BACKGROUND: Recent data indicate that cardiac surgery with cardiopulmonary bypass (CPB) results in an imbalance of T-helper cell subsets towards the anti-inflammatory pathway mediating humoral immune response. However, little is known about immunoglobulin levels as an important part of humoral immune response after CPB. Therefore, the objectives of this study were 1) to elucidate the effects of CPB on perioperative immunoglobulin levels, and 2) to find out if alterations in lymphocyte subsets are related to these findings. METHODS: Blood samples from 83 patients undergoing elective cardiac operation were taken preoperatively (d0), on the first (d1), third (d3), and fifth day (d5) after operation. Levels of immunoglobulin (Ig) E, IgM, and IgG, including the subclasses IgG 1 - 4, were measured. IgG2/IgE-ratio was used as indicator for TH1/TH2 shifting, and production of tetanus antibodies (AB) was investigated as an in vivo parameter of humoral immune reaction. The number and percentage of T- and B-lymphocyte subsets were assessed in a subgroup of 50 patients to answer the second question. RESULTS: Clinically, no mortality or major morbidity were observed. IgE levels did not change until d3 and increased significantly on d5. In contrast, both IgG and IgM levels decreased significantly on d1. While IgM returned to baseline (BL) on d5, IgG levels remained below BL until d5. IgG2/IgE-ratio decreased significantly on d1, reached its nadir on d3 and remained depressed until d5. The number of T-lymphocytes decreased on d1 as well as the number of B-cells. T-cells returned to BL on d5, B-cells on d3. However, while the percentage of T-cells decreased on d1, the percentage of B-cells increased. The percentage of T-cells returned to BL on d3, and B-cell percentage returned to BL on d5. Tetanus AB production did not change until d5 when it increased significantly. CONCLUSIONS: 1) Increase of IgE and tetanus AB production indicate that humoral immune response is not affected by CPB, but possibly even enhanced. The relative increase of B-cells is in line with this hypothesis. 2) Postoperative changes in immunoglobulin levels provide further evidence for a TH1/TH2-shifting. 3) The transient deficit in IgM-and IgG levels did not result in clinically adverse events. Thus, therapeutic intervention appears not to be required.

Adult↗

Effect of controlled antigenic stimulation on lymphocyte subsets in pigs and pig fetuses.

The effect of controlled antigenic stimulation in immunologically virgin organisms, i.e. pig fetuses treated with NDCM (Nocardia delipidated cell mitogen) and germ-free (GF) piglets associated with a non-pathogenic E. coli 086, on peripheral blood lymphocyte subsets defined by the expression of CD5 and CD8 was studied by double color flow cytometry. Stimulation of both fetuses and GF piglets increased the frequency of CD8low+ lymphocytes. A prominent subset of CD5-CD8low+NK cells was present in GF and E. coli associated piglets and their frequency was slightly higher in E. coli associated animals. The most pronounced difference between stimulated and non-stimulated animals was in a relative proportion of an ill-defined lymphocyte subset with an unusual CD5low+CD8low+ expression. Both NDCM injection into fetal blood circulation and association of GF piglets with E. coli resulted in a marked increase of frequency of CD5low+CD8low+ lymphocytes in peripheral blood.

Adjuvants, Immunologic↗