ABSTRACT
Production of autoantibodies is one of the main features of primary Sjögren's syndrome (pSS). Long-lived plasma cells (PC) can produce autoantibodies for prolonged period of times without being affected by immunosuppressive therapies. As of today, little is known about the long-lived PC subset and their contribution to autoimmunity. We have characterized the phenotypic and migratory properties of peripheral blood PC isolated from pSS patients (grouped by focus score, FS) and compared them to PC from rheumatoid arthritis (RA) patients and normal non-autoimmune subjects. We observed two populations of PC in all study groups, CD19+ PC and CD19- PC. Interestingly, the CD19- PC subset was most prominent in autoimmune patients (pSS and RA) compared to normal controls. Further investigation of the PC phenotype revealed that a high percentage of both CD19+ and CD19- PC isolated from pSS and RA patients did not express the CD27 marker, which is normally highly expressed on all types of PC. Differences in the expression of markers such as IgM, IgG, CD95 and CXCR3 in the group with high FS compared to FS = 1, underscore the heterogeneity of pSS patient group and demonstrate that phenotypic pattern of circulating PC associates with the severity of inflammation in the salivary glands of these patients. Our migration experiments show that addition of CXCL12 to PC in vitro, do not alter the migration potential of PC in any group tested. However, we observed an overall higher spontaneous migration of PC from pSS compared to both RA and normal controls.
Subject(s)
Arthritis, Rheumatoid/immunology , Autoantibodies/blood , Plasma Cells/immunology , Sjogren's Syndrome/immunology , ADP-ribosyl Cyclase 1/blood , Antigens, CD19/blood , Arthritis, Rheumatoid/blood , Cell Movement/immunology , Female , Flow Cytometry , Humans , Male , Middle Aged , Receptors, CXCR3/blood , Receptors, CXCR4/blood , Sjogren's Syndrome/blood , Syndecan-1/bloodABSTRACT
Vaccination provides the most effective method of limiting the impact of influenza. Inactivated influenza vaccines are available in three formulations and more information needs to be generated on how antigen presented in different vaccine formulations influences the subsequent immune response. In the present study, we have investigated the effect of two different influenza vaccine formulations on the resulting antibody and cytokine responses and compared these responses with influenza infection. Mice were vaccinated intramuscularly with one or two doses of whole or split virus vaccine or alternatively intranasally infected with influenza virus. Lymphocytes were isolated from spleen cells and stimulated in vitro for 24 or 72 h for analysis of cytokine profile at the gene expression and at the protein level. Additionally, whole blood was collected and the serum antibody response investigated by haemagglutination inhibition (HI) and enzyme-linked immunosorbent assay (ELISA). We found that one dose of whole virus vaccine induced higher antibody and cytokine responses and thus was more immunogenic in unprimed mice than split virus vaccine. Whole virus vaccine induced a strong IFN-gamma (type 1) immune response after one dose of vaccine and a more mixed cytokine response after the second dose. Split virus vaccine induced a type 2 response, particularly after two vaccine doses. Our results show that two doses of vaccine (both vaccine formulation) were more effective in induction of Th2 type of cytokines and thus indicate that both the formulation and also the number of vaccine doses substantially influences the magnitude and quality of the immune response.
Subject(s)
Antibodies, Viral/blood , Cytokines/metabolism , Influenza A Virus, H3N2 Subtype/immunology , Influenza Vaccines/immunology , RNA, Messenger/metabolism , Vaccination/methods , Animals , Antibody-Producing Cells/immunology , Dose-Response Relationship, Immunologic , Female , Gene Expression , Hemagglutinins/pharmacology , Mice , Mice, Inbred BALB CABSTRACT
A gas-chromatographic method for isolation and determination of blood chlorpropamide, described originally in literature [10], was modified and its usefulness for clinical and pharmacological purposes evaluated by testing fluctuations in blood chlorpropamide concentration with time in healthy subjects (control group) and patients with type II diabetes mellitus, in relation to glycemia and insulinemia (IRI). In this respect the effects of a single dose and of prolonged chlorpropamide therapy were studied. Large individual variability in the time-related concentration curves and no statistically significant correlation between chlorpropamide, insulin and glycemia were noted.
Subject(s)
Chlorpropamide/blood , Chromatography, Gas/methods , Adult , Blood Glucose/analysis , Female , Humans , Insulin/blood , Male , Middle AgedSubject(s)
Hydrazines/metabolism , Polarography/methods , Todralazine/metabolism , Animals , Female , In Vitro Techniques , Male , Rabbits , Rats , Todralazine/blood , Todralazine/urineABSTRACT
Noxyptyline, i.e. 5-(2-dimethylaminoethyloxyimine)-5H-dibenzo[a,b] cyclohepta-1,4-diene hydrochloride, is an antidepressant. A new, direct method for its determination in substance and in tablets by means of gas chromatography has been developed. The results were compared with those of the spectrophotometric method, and the systematic errors(coefficient of variation) were 2.19% and 2.49%, respectively. Appropriate conditions were developed for the extraction of noxyptyline from plasma and urine, and the gas chromatographic method was applied for its determination. Within the concentration range 0.5-10 microgram/cm3, the systematic error after extraction from plasma was 4.61%, and after extraction from urine it was 2.08%. The recovery from plasma was 71.12 +/- 8%, and from urine it was 90.94 +/- 1.5%.
