Positive Peripheral Blood Lymphocytes Research Articles

Dietary encapsulated essential oil mixture influence on apparent nutrient digestibility, serum metabolic profile, lymphocyte histochemistry and intestinal morphology of laying hens.

ObjectiveThe study aimed to evaluate the effects of a mixture of encapsulated essential oils (EOs) addition on nutrient digestion, serum biochemical parameters, peripheral blood alpha-naphthyl acetate esterase (ANAE), and acid phosphatase (ACP-ase) positive lymphocyte ratios and intestinal morphology in laying hens.MethodsA total of 320 laying hens of 48-wk-old were randomly allotted into 4 treatment groups with 10 replicates of 8 birds in each replicate. The birds were fed a basal diet (control) or the diet added with mixture of EOs (which consist of eugenol, nerolidol, piperine, thymol, linalool, and geraniol) at 50, 100, and 200 mg/kg for period of 84 days.ResultsThe addition of EOs at 100 or 200 mg/kg increased the dry matter, organic matter, and crude protein digestion as compared to control. The addition of all doses of EOs did not affect serum gamma glutamyl transferase, alanine aminotransferase, and P but increased serum asparate aminotransferase (AST) concentration. The addition of 200 mg/kg EOs increased serum creatinine, while 100 mg/kg decreased Ca concentration. The addition of 100 and 200 mg/kg EOs generally improved ANAE and ACP-ase positive peripheral blood lymphocyte ratios and intestinal morphology.ConclusionIt can be concluded that, the addition of 100 or 200 mg/kg encapsulated EOs generally increased apparent nutrient digestion and serum AST concentration, improved ANAE and ACP-ase positive peripheral blood lymphocytes and intestinal morphology in laying hens.

Effects of paclitaxel and resveratrol on blood characteristics in rabbits

ABSTRACTWe investigated the adverse effects of paclitaxel (PAC) on blood characteristics including the percentage of alpha naphthyl acetate esterase (ANAE)-positive peripheral blood lymphocytes, white cell count, number of neutrophil nuclear lobe and, erythrocyte diameter, and the protective effects of resveratrol (RES) on these characteristics. Male New Zealand rabbits were assigned to four groups. The control group (C) was given 40 ml normal saline, the PAC group was given 5 mg/kg PAC in 40 ml normal saline, the RES group was given 4 mg/kg RES in 40 ml normal saline, and the PAC + RES group was given 5 mg/kg PAC + 4 mg/kg RES in 40 ml normal saline. All injections were performed intravenously once/week for 8 weeks. PAC caused an increased total percentage of lymphocytes and monocytes, which was associated with neutropenia. The PAC group showed a right shift in the lobulation curve of neutrophil nuclei together with a decreased percentage of ANAE-positive lymphocytes. RES reduced the percentage of ANAE-positive lymphocytes, slightly increased the percentage of neutrophil leukocytes and reduced the total lymphocyte count. Administration of RES combined with PAC prevented, while PAC induced, neutropenia and lymphocytosis, and increased the percentage of ANAE-positive lymphocytes.

Cytochemistry of peripheral blood leukocytes in thoroughbred foals

We attempted to characterize the cytochemical staining patterns of leukocytes and to determine the percentages of alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACPase) positive lymphocytes in peripheral blood of thoroughbred foals at different ages. Blood samples were obtained from the jugular veins of 60 healthy thoroughbred foals, 1 day, 3 days and 1 year old. Each age group included 10 male and 10 female animals. Peroxidase (PO) activity was detected in neutrophils, eosinophils and monocytes. Lymphocytes were negative for PO staining. Periodic acid-Schiff (PAS) staining was observed in neutrophils and monocytes; eosinophils were negative. The majority of lymphocytes were negative, but a few cells showed granular PAS positivity. Monocytes were strongly positive for ANAE and ACPase, and the enzymatic reaction was common in lymphocytes. Neutrophils showed a weakly positive reaction for ANAE and ACPase. Eosinophil granules were negative or weakly positive for ANAE and usually negative for ACP, but intergranular areas were positive. Mean ANAE positive peripheral blood lymphocytes (PBL) were 67.70, 73.10 and 49.20% in females; 64.00, 70.53, and 50.60% in males 1 day, 3 days, and 1 year old, respectively. ACPase positive PBL were 27.33, 32.83, and 37.40% in females; 29.67, 31.67, and 38.40% in males 1 day, 3 days, and 1 year old, respectively. For both enzymes, the differences between mean values for the genders were not statistically significant, but significant differences were found with regard to age. We provide comparative hematological data as a guide for identifying blood cells in thoroughbred foals.

Clinical immunology Th1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis

ENWEndNote BIBJabRef, Mendeley RISPapers, Reference Manager, RefWorks, Zotero AMA Almogren A, Adam M, Shakoor Z, Gadelrab M, Musa H. Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis. Central European Journal of Immunology. 2013;38(1):100-106. doi:10.5114/ceji.2013.34365. APA Almogren, A., Adam, M., Shakoor, Z., Gadelrab, M., & Musa, H. (2013). Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis. Central European Journal of Immunology, 38(1), 100-106. https://doi.org/10.5114/ceji.2013.34365 Chicago Almogren, Adel, Mustafa Hussein Adam, Zahid Shakoor, Mohammad Osman Gadelrab, and Hassan Abdulaziz Musa. 2013. "Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis". Central European Journal of Immunology 38 (1): 100-106. doi:10.5114/ceji.2013.34365. Harvard Almogren, A., Adam, M., Shakoor, Z., Gadelrab, M., and Musa, H. (2013). Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis. Central European Journal of Immunology, 38(1), pp.100-106. https://doi.org/10.5114/ceji.2013.34365 MLA Almogren, Adel et al. "Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis." Central European Journal of Immunology, vol. 38, no. 1, 2013, pp. 100-106. doi:10.5114/ceji.2013.34365. Vancouver Almogren A, Adam M, Shakoor Z, Gadelrab M, Musa H. Clinical immunologyTh1 and Th2 cytokine profile of CD4 and CD8 positive peripheral blood lymphocytes in nickel contact dermatitis. Central European Journal of Immunology. 2013;38(1):100-106. doi:10.5114/ceji.2013.34365.

Hematology and enzyme histochemistry of peripheral blood lymphocytes in domestic pigeon (Columba livia f. domestica)

The purpose of our study was to determine the percentages of alpha-naphthyl acetate esterase (ANAE)-positive and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL) in both sexes of one-year-old domestic pigeons (Columba livia f. domestica). Blood samples obtained from 12 healthy domestic pigeons were used. The mean percentages of ANAE-positive PBL for females and males were 47.8% and 48.8%, respectively, whereas the mean percentages of ACP-positive PBL were 67.5% and 68.6%, respectively. The proportions of PBL were 49.3% and 48.6% in males and females, respectively.

442 Coincidence of Celiac Disease and Gluten Allergy

BackgroundThe type I or IV of hypersensitivity reactions according to Gell and Coombs classification may be responsible for clinical symptoms observed after ingestion of gluten - containing products. The mechanisms of these reactions are either IgE-dependent or IgE-independent. Celiac disease based on IgE-independent mechanism is classified as gluten hypersensitivity. Clinical manifestation of celiac disease and gluten allergy is often similar. Correct diagnosis of this disease is particularly important due to the different long-term therapeutic procedures. We would like to asses of the incidence of celiac disease in children with gluten allergy.MethodsThe study involved 50 children with abdominal pain, chronic diarrhea, recurrent respiratory and ears inflammation and skin lesions - patients of the Immunological and Gastroenterology Outpatient Clinic of Institute of Mother and Child. The allergy to gluten was confirmed on the basis of positive peripheral blood lymphocytes blast transformation test and detection of allergen-specific IgE antibodies to gluten (f79). In all children plasma concentration of immunoglobulin classes A, G M and IgA or IgG antibodies against tissue transglutaminase (tTGA) were measured.ResultsIn children on the study group the type IV of hypersensitivity reaction to gluten was diagnosed. In 3 children specific IgE antibodies to gluten was also confirmed (f79 - I type hypersensitivity). Anti-tissue transglutaminase antibodies both IgA and IgG were detected in 2 children in whom the concentration of IgA and IgG in serum remained within normal range for age. In these children celiac disease was confirmed by jejunal biopsy.ConclusionsThe predominant frequency of type IV of hypersensitivity reactions in children in response to the gluten antigen should be taken into account in diagnosis of food allergy.In children diagnosed with gluten allergy the test for celiac disease should be performed.

Investigation of α-naphthyl acetate esterase and acid phosphatase in the peripheral blood leukocytes of greyhounds

The purpose of our study was to determine the percentages of α-naphthyl acetate esterase (ANAE)- and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL), the presence of the ANAE and ACP enzymes in leukocytes, and the proportion of PBL in greyhounds. Peripheral blood samples were collected from the cephalic antebrachial vein of 14 (7 animals of each sex) healthy 1-2-year-old greyhounds. Mean percentages of ANAE-positive PBL were found to be 73.29 ± 0.95% in female and 74.29 ± 2.21% in male dogs. The difference between mean values of the genders was not statistically significant. The ACP values were 36.00 ± 2.94% for females and 33.57 ± 2.15% for males. No significant differences were found with regard to gender. For both enzymes, although monocytes and eosinophilic granulocytes displayed a positive reaction, neutrophils gave negative reactions. The proportion of PBL was 36.29 ± 5.31% and 33.00 ± 2.38 % in female and male dogs, respectively. The differences were not significant.

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.

Hematology and Enzyme Histochemistry of the Peripheral Blood Leucocytes in Rock Partridges (Alectoris graeca)

The purpose of this study was to determine the percentages of alpha-naphthyl acetate esterase (ANAE)-positive and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL) and the percentages of leukocytes in the rock partridge at different ages. Blood samples obtained from the vena basilica of 18 healthy rock partridges (Alectoris graeca) at 1 d, 5 wk, and 12 wk of age were used. Mean percentages of ANAE-positive PBL for the 3 age groups were 37, 29.83, and 47.83% for 1 d, 5 wk, and 12 wk of age, respectively. Heterophils also gave ANAE-positive reactions. Mean percentages of ACP-positive PBL for the 3 age groups were 70, 81, and 86.1% for 1 d, 5 wk, and 12 wk of age, respectively. Although monocytes showed a diffuse granular staining pattern, heterophils displayed a weak positive reaction for ACP. Thrombocytes showed a small granular staining pattern. This study contributes by broadening the hematological research on avian species and provides a guideline for identifying blood cells in the rock partridge.

間質性肺炎発症から後天性免疫不全症候群と判明した2症例

Case 1: A 35-year-old man admitted for fever and respiratory failure during several weeks was found in chest computed tomography (CT) to have interstitial pneumonia, and the plasma beta-D-glucan level indicated Pneumocystis jiverocii pneumonia. Psoriasis from second-stage syphilis raised the suspicion of HIV infection. Serum anti-HIV-1 antibody proved positive and CD4-positive lymphocytes in peripheral blood were 18/microL. The man died despite treatment. Autopsy confirmed P. jiverocii pneumonia. Case 2: A 28-year-old man seen for a fever and respiratory failure was found in chest CT to have mild interstitial pneumonia. We checked for hypersensitivity pneumonitis, Mycoplasma pneumoniae pneumonia, etc. The plasma beta-D-glucan level indicated possible P. jirovecii pneumonia and immunodeficiency. Serum anti-HIV-1 antibody proved positive and CD4-positive lymphocytes in peripheral blood were 34/microL. The man was treated successfully, using trimethoprim with sulfamethoxazole for his interstitial pneumonia. His clinical symptoms were compatible with P. jirovecii pneumonia. P. jirovecii pneumonia with AIDS may present with more subacute or subtle symptoms than other immunosuppressive diseases, making it difficult to diagnose. Medical professionals should thus make it a point to familiarize themselves with AIDS prevention.

<I>Bifidobacterium pseudolongum</I> JBP01株含有腸溶性カプセルを投与された犬における免疫応答

This study investigated immune responses in 5 normal male beagle ranging from 3 to 6 years old orally given an enteric capsulate preparation containing Bifidobacterium pseudolongum (JPB01). After 30 days, the number and ratio of CD4-positive lymphocytes, the number of CD3-positive lymphocytes, and the ratio of CD4/CD8 positive lymphocytes in peripheral-blood were significantly higher in all 5 dogs than those before inoculation. The number of CD25-positive lymphocytes after the stimulation by mitogen irritants (PHA, ConA and PWM) had increased significantly. No significant difference in granulocyte phagocytosis was observed between pre and post-inoculation. These data suggest that B. pseudolongum had immunepotentialized lymphocytes.

Transplantation of dermal multipotent cells promotes the hematopoietic recovery in sublethally irradiated rats.

Our previous study indicated that dermal multipotent cells with the differentiation capacity to form cells with the phenotypic properties of osteocytes, adipocytes, chondrocytes, and neurons in specific inducing media could be isolated from the enzymatically dissociated dermal cells of newborn rats by their adherence to culture dish plastic. We have also observed that the systemic transplantation of dermal multipotent cells could not repopulate the hematopoietic system in lethally irradiated rats. In this paper, we found that a transplantation of plastic-adherent dermal multipotent cells into sublethally irradiated rats led to a significant increase of white blood cells in peripheral blood, nucleated cells, CFU-GM, and CFU-F colonies in bone marrow. FISH analysis, using a Y-chromosome specific probe, showed that dermal multipotent cells could engraft into bone marrow in recipients. Flow cytometry (FACS) analysis also showed that the proportion of CD2 and CD25 positive lymphocytes in peripheral blood did not change significantly in two weeks after transplantation. By these results, we infer that dermal multipotent cells may represent an alternative origin of mesenchymal stem cells to restore marrow microenvironment and promote the survival, engraftment, and proliferation of hematopoietic cells.

Immunocytochemical identification of CD5 positive peripheral blood lymphocytes in four marsupial species

Immunocytochemical analysis of peripheral blood mononuclear cells was undertaken using a streptavidin biotin–horseradish peroxidase method to detect CD5 positive lymphocytes from the blood of several marsupial species. A monoclonal antibody raised to a conserved peptide sequence of the human CD5 antigen positively labelled lymphocytes in freshly isolated peripheral blood mononuclear cells of the tammar wallaby (Macropus eugenii), the long-footed potoroo (Potorous longipes), the long-nosed potoroo (Potorous tridactylus) and the rufous hare-wallaby (Lagorchestes hirsutus). A polyclonal anti-CD3 antibody also positively labelled circulating lymphocytes from the tammar wallaby. Whereas previous studies using flow cytometry reported labelling of T cells in koala lymphocyte preparations using a polyclonal anti-CD3 antibody, there have been no other reports of marsupial blood immunophenotyping. The current study extends the known applications of monoclonal anti-CD5 and polyclonal anti-CD3 antibodies to blood lymphocytes of small wallaby species using an immunocytochemical slide technique that is simple, can be processed within a day and requires no dedicated large equipment.

Eating disorders: a role for dipeptidyl peptidase IV in nutritional control

Dipeptidyl peptidase IV (DPP IV), a serine protease with broad tissue distribution and known activity in serum, has been postulated to modulate nutrition control by modification or inactivation of peptide hormones operating in the enteroinsular axis. We hypothesized that changes of DPP IV activity in serum are related to the nutrition status of patients with eating disorders. Serum DPP IV activity was measured in 52 patients (28 with anorexia nervosa and 24 with bulimia nervosa) in four consecutive weekly analyses. Simultaneously, the number of CD26 (DPP IV)-positive peripheral blood lymphocytes was counted. The same analyses were carried out in 28 healthy female volunteers. In week 1 and throughout the observation period, DPP IV activity in the sera of patients with anorexia nervosa and, to a lesser extent, those with bulimia nervosa was elevated in comparison to that of healthy controls (week 1: means = 92.8 U/L for anorexia-nervosa patients and 89.3 U/L for bulimia-nervosa patients versus 74.7 U/L for healthy control subjects, P = 0.014; weeks 1–4: 91.8 U/L for anorexia-nervosa patients and 86.2 U/L for bulimia-nervosa patients versus 77.6 U/L for healthy controls, P < 0.001). We assume that the increase in DPP IV serum activity will increase the turnover of distinct peptide hormones with known effects on nutrition control and susceptibility to degradation by DPP IV. The potential impact of an increase in DPP IV activity in serum on satiety and nutrition control contributes to previously reported implications for immune function.

Dipeptidyl Peptidase IV (DP IV, CD26) in Patients with Inflammatory Bowel Disease

Background: Dipeptidyl peptidase IV (DP IV, CD26), a serine protease with broad tissue distribution and known activity in serum, participates in T cell activation and promotes a Th1 cytokine response, a function in part attributable to its enzymatic activity. We hypothesized that the activity of DP IV in serum and expression of CD26/DP IV in lymphocytes may be altered in patients with inflammatory bowel disease (IBD). Methods: Serum DP IV activity and CD26 (DP IV)-positive peripheral blood lymphocytes were measured in 110 patients with IBD (Crohn disease (CD): n = 63, ulcerative colitis (UC): n = 47). Additionally, T cell activation antigens (CD25, CD95) and costimulatory molecules (CD28) were evaluated. The same analyses were carried out in healthy volunteers (HC, n = 28). Thirty-nine patients with CD and 28 patients with UC were reassessed 3-6 months after the first visit. Results:

Expression of Fas antigen (CD95) in peripheral blood lymphocytes and in liver-infiltrating, cytotoxic lymphocytes in patients with hepatocellular carcinoma.

Fas-expressing cytotoxic T lymphocytes (CTLs) are important antitumor immune effector cells in patients with hepatocellular carcinoma (HCC). The role of transforming growth factor beta 1 (TGF-beta1) in modulating the expression of Fas by CTLs is not known in HCC. The objectives of this study were to characterize the expression of Fas by CTLs and natural killer (NK) cells among peripheral blood lymphocytes (PBLs) and tumor-infiltrating lymphocytes (TILs) in patients with HCC and to correlate the association, if any, with serum TGF-beta1 levels. PBLs from 18 patients with HCC and TILs from 5 HCC liver specimens were isolated, and Fas expression was analyzed by three-color flow cytometry. The results were compared with results from normal control volunteers (n = 19 individuals). Serum TGF-beta1 levels in patients with HCC were measured by enzyme-linked immunosorbent assay. The median percentage of Fas expression by CD3 positive T cells was significantly higher in patients with HCC compared with normal controls (54.37% vs. 32.03%, respectively; P = 0.0036), and this was attributable solely to Fas expression by CD4 positive PBLs (54.46% vs. 34.90%, respectively; P = 0.0234). In contrast, Fas expression was significantly higher in all the subtypes of TILs (CD3 positive, CD4 positive, CD8 positive, NK cells, and natural T cells) compared with controls (all P values were < 0.001). Tumor size was inversely proportional to the TGF-beta1 levels (correlation coefficient [r] = -0.725; P < 0.0001), which were correlated inversely with Fas expression by CD4 positive PBLs (r = -0.516; P = 0.01). In patients with HCC, TILs exhibit significantly increased expression of Fas compared with PBLs that may enhance their susceptibility to apoptotic mechanisms. Larger tumors were associated with lower serum TGFbeta1 levels, and this was correlated with greater Fas expression by CD4 positive PBLs.

Fine-tuning the EBV+ hu-PBL-SCID xenogeneic chimera model usingIn Vivo superinfection

Our purpose was to establish a reproducible xenogeneic chimera model to observe tumors similar to the well-known human posttransplant lymphoproliferative disease (LPD). First we followed the original protocol injecting Epstein-Barr virus positive (EBV+) human peripheral blood lymphocytes (PBL) intraperitoneally into immunodeficient (SCID) mice. Human cells showed T cell phenotype in majority one week after the transfer, whereas one month later a shift towards B cell phenotype was evident according to immunohistochemical and flow cytometric analysis. At this stage the intraperitoneal mass of cells suggested a biologically malignant behaviour infiltrating the liver and the spleen of the host animal. Immunohistochemistry indicated proliferating human lymphatic cells expressing EBV associated proteins and characteristic patterns of invasion within the affected organs. Eventually LPD was lethal to the host animals in 46-67 days. However, the microscopic appearance of experimental LPD was different from the human haemopoietic malignancies: the basic structures of lymphatic organs were preserved and the human T and B cells repopulated the normally T and B dependent areas in mice. The phenotypes of the proliferating cells were human and characteristic for the mature T- and B-lymphocytes. No dominant clone developed during in vitroculturing of the biologically invasive mass of cells removed from the tumor-bearing mice. The results of microscopical, immunological, and flow cytometrical analysis suggested a mature but uncontrolled proliferation of human lymphocytes in SCID mice. The original method for the induction of post-transplant LPD in SCID mice was modified in our further experiments to standardise the experimental technique increasing the efficiency of B cell proliferation and the reducing the number of unspecific factors. Subsequent in vivo EBV superinfection was carried out after the intraperitoneal transfer of a reduced quantity of human PBL from different donors. The same disease developed in our modified chimera model as by the use of original protocol except for some valuable differences. All hosts developed LPD regardless the significantly reduced amount of transplanted PBL and it was lethal in a shorter period of time (41-43 days) compared to the original model. The decreased quantity of transplanted human lymphatic cells was formerly insufficient using the original protocol. Therefore this modified and standardised protocol can lead to a more predictable and reproducible model allowing us to examine fine details of posttransplant lympho-proliferative disease.

Alterations in expression and in serum activity of dipeptidyl peptidase IV (DPP IV, CD26) in patients with hyporectic eating disorders.

The notion that patients with eating disorders maintain a functional immunosurveillance in spite of severe malnutrition has attracted researchers for years. Dipeptidyl peptidase IV (DPP IV), a serine protease with broad tissue distribution and known activity in serum, operates in the cascade of immune responses. Membrane-bound DPP IV expressed on lymphocytes, also known as the leucocyte antigen CD26, is considered to participate in T-cell activation. We hypothesized that the activity of DPP IV in serum and expression of CD26 in lymphocytes may be altered in patients with eating disorders. Serum DPP IV activity and the number of CD26 (DPP IV)-positive peripheral blood lymphocytes were measured in 34 patients [anorexia nervosa (AN): n = 11, bulimia (B): n = 23] in four consecutive weekly analyses. In addition, the expression of CD25 (interleukin-2 receptor alpha chain) was evaluated to estimate the degree of T-cell activation. The same analyses were carried out in healthy female volunteers (HC, n = 20). CD2-CD26-positive cells were reduced in patients compared with healthy controls [mean 40.2% (AN) and 41.1% (B) versus 47.4% (HC), P < 0.01], while the DPP IV activity in serum was elevated [mean 108.4 U/l (AN) versus 91.1 U/l (B) and 80.3 U/l (HC), P < 0.01]. The potential implications of our observations on, and beyond, immune function are discussed.

Increased levels of IL-5 positive peripheral blood eosinophils and lymphocytes in mild asthmatics after allergen inhalation provocation.

In allergic inflammation eosinophils and TH2-like lymphocytes are supposed to be the major effector cells and considered to contribute as cellular source of the key cytokine interleukin (IL)-5. The purpose of this study was to enable detection of IL-5 containing leucocytes and to investigate whether the number of these cells in the blood circulation differed between healthy and asthmatics before and after allergen provocation. The distribution of intracellular IL-5 in human peripheral blood eosinophils (PBE) and lymphocytes (PBL) has been investigated using fixation and cell membrane permeabilization with octyl-glucopyranoside, the FOG-method, and flow cytometry. The intracellular staining was performed on leucocytes without any prior purification and in vitro stimulation. The specificity of IL-5 binding to intracellular compartment of both PBE and PBL was confirmed by complete inhibition with human recombinant IL-5. Preformed intracellular IL-5 was detected in the main population of PBE (> 70%) in both healthy individuals and asymptomatic patients. Moreover, preformed intracellular IL-5 was also detected in 4.8% and 2.4% of PBL from healthy individuals and asymptomatic patients, respectively. There was a correlation between the absolute number of PBE and IL-5 positive PBE. In patients with pollen-related asthma, the number of IL-5 positive PBE and PBL increased significantly 24 h after an allergen inhalation provocation (P < 0.05). In the healthy control group no differences regarding IL-5 positive PBE and PBL were obtained pre- and post-allergen challenge. In patients with mild allergic asthma, but not in healthy individuals, allergen provocation induces an increased absolute number of IL-5 positive PBE and PBL. The reason for the relatively high number of IL-5 positive PBL is unclear, but a plausible explanation might be that other lymphocyte subsets besides CD4+ TH2 can produce IL-5. However, enumeration of IL-5 positive leucocytes may be used as an activity marker and also be a useful tool in monitoring the inflammation in asthma.

Th2 subset dominance among peripheral blood T lymphocytes in patients with digestive cancers

BACKGROUND: Two types of helper T cells (Th), which are categorized as Th1 and Th2 on the basis of cytokine production, have been reported. Th1 cells produce interleukin (IL)-2 and interferon (IFN)-γ, while Th2 cells secrete IL-4, IL-6, and IL-10. We assessed the intracellular cytokine profiles of CD3/CD4 positive lymphocytes (CD4 + T-cells) in peripheral blood in patients with digestive cancers. METHODS: Peripheral blood samples were collected from 50 patients with digestive cancers and 35 healthy volunteers. The proportions of CD4 + T-cells producing intracellular cytokines were determined using flow cytometry. RESULTS: The percentages (mean ± SD) of CD4 + T-cells producing IL-4, IL-6, and IL-10 in the cancer group (73.9% ± 13.0%, 73.0% ± 16.6%, and 58.0% ± 21.0%, respectively) were significantly higher than in the healthy group (37.4% ± 12.4%, 37.8% ± 13.5%, and 34.0% ± 14.1%, respectively; P <0.01). Proportions of CD4 + T-cells producing IL-4, IL-6, and IL-10 in 10 patients undergoing curative resection had decreased significantly 1 month after surgery ( P <0.01). No significant difference was noted between groups in the percentages of CD4 + T-cells producing IFN-γ. CONCLUSIONS: Th2-dominant status develops in cancer patients. Such lymphocyte evaluations could find applications in diagnosis and therapeutic monitoring of cancer patients.