Antibody immunoglobulin G1 and immunoglobulin G2a responses against some cystic fluid proteins of Cysticercus bovis in Balb/c mice

Background and Aim: Immunoglobulin (Ig) G1 and IgG2a are the surrogate markers respectively for humoral and cellular immune responses of hosts against antigens including cystic fluid proteins of Cysticercus bovis. A study was conducted to investigate the IgG1 and IgG2a responses of Balb/c mice against some individual cystic fluid proteins of C. bovis in an effort to determine the roles of each protein in inducing the humoral and cellular immune responses in host. Materials and Methods: Individual p71, p31, and p14 proteins of C. bovis were purified by separation of the proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and elution of individual proteins from the gel. Six female Balb/c mice were immunized 4 times at 10-day intervals with the crude cystic fluid proteins, and sera were collected for the measurement of IgG1 and IgG2a levels against the individual proteins. Sera samples collected before the first immunization were used as negative antibody control, sera samples collected after the fourth immunization were used as positive antibody control, and crude cystic fluid protein was used as positive antigen control. Results: All immunized mice were immune to p71, p31, p14, and crude cystic fluid proteins of C. bovis. The crude cystic fluid proteins of C. bovis induced a higher IgG2a than IgG1 level following the first and the second immunizations but switched into a higher IgG1 than IgG2a level following the fourth immunization. Protein 71 kDa (p71) induced a higher IgG2a than IgG1 level following the fourth immunization. In contrast, p14 induced a higher IgG1 than IgG2a level following the fourth immunization. Low and balance IgG1 and IgG2a levels against p31 were observed following the first to the fourth immunizations. Conclusion: Using IgG1 and IgG2a levels as the surrogate markers, it appears that cystic fluid antigens of C. bovis induce both humoral and cellular immune responses in Balb/c mice. The p71 appears to be a better inducer of cellular immune response, whereas p14 is a better inducer of humoral immune response of mice.

Study of IgG Subclass of the Patients with Immuno-Related Hemocytopenia

Objective: To investigate the immunoglobulin G (IgG)sublass on the membrane of bone marrow(BM) hematopoietic cells and IgG levels in BM supernatant of patients with immuno-related hemtocytopenia(IRH). Method: Newly diagnosed IRH patients and ICUS patients were enrolled in this study, as well as healthy donors as controls. The categories of auto-antibodies(IgM and IgG) on the membrane of BM hemmatopoietic cells(CD15+,CD34+ and GlyCoA+ cells)were assayed by Flow cytometry(FCM). The categories of auto-antibodies(IgG1 and IgG3) on the membrane of BM hemmatopoietic cells were assayed by FCM.BM nucleated cells antibodies (IgG1AIgG2AIgG3 and IgG4)were detected by immunohistochemical staining. The IgG1AIgG2AIgG3 and IgG4 levels in BM supernatant were detected by Cytometric Bead Array (CBA)GThe ratio of CD5+CD19+/CD19+ in blood were tested by FCMGThe mRNA expression of IgG1 in peripheral blood were detected by RT-PCR. Result: The positive rate of antibodies (IgG) on BM hemmatopoietic cells of IRH patients was 58.69(27/46), which was significantly higher than that in ICUS group(0%) and healthy control group(0%). The positive rate of IgG1 on CD15+ cells was (3.13±1.80)% in IRH group, which was significantly higher than that in healthy control group. The positive rate of IgG3 on CD15+ cells was(2.73±2.09)% in IRH group, which was significantly higher than that in healthy control group. The positive rate of IgG1 on GlyCoA+ cells was( 3.97±2.21)% in IRH group,which was significantly higher than that in healthy control group. The positive rate of IgG3 on GlyCoA+ cells was (2.24±1.82)% in IRH group,which was significantly higher than that in healthy control group. IgG subclass(IgG1AIgG2AIgG3 and IgG4)on BM nucleated cells were detected by Immunohistochemistry. 11/15 IRH patients' IgG1 positive cells≥3(count 100 bone marrow nucleated cells). There were rarely IgG1+ cells in ICUS patients and healthy donors' bone marrow smears. IgG2,IgG3 and IgG4 positive BM nucleated cells were rarely detected in three groups by immunohistochemistry. The IgG1 level in IRH group was (6998±2498)mg/L which was higher than remission group (4702±1235)mg/L, healthy control group (5613±1640) mg/L and ICUS group (5166±1761) mg/L, P<0.05. The IgG3 level in IRH group was (637±252)mg/Lwhich was higher than remission group(415±196)mg/L,healthy control group (422±212) mg/L and ICUS group (485±197) mg/L, P<0.05. IgG2 levels in four groups were no statistical difference. IgG4 levels in these four groups were no statistical difference. The total IgG level in IRH group was (10903±2576)mg/L which was higher than remission group(8468±1957)mg/L, P<0.05. The ratio of CD5+CD19+/CD19+ in IRH group was(23.34±9.36)% which were significance higher than healthy control group(13.67±5.10)% and ICUS group(12.40±3.63)% (P<0.01). IgG1 level had a positive correlation with the proportion of CD5+ B lymphocytes(r=0.392, P =0.043). There was an inverse correlation between the IgG1 level and the count of neutrophil (r = -0.459, P = 0.016) in IRH patients. There was an inverse correlation between the level of IgG3 and hemoglobin(Hb) (r = -0.389, P = 0.045) in IRH patients. In IRH group, the mRNA expression of IgG1 in peripheral blood is 5.27±4.83 which is higher than healthy control 1.58±1.50, P<0.05. Conclusion: There are auto-antibodies IgG on the membrane of BM hematopoietic cells of IRH patients. IgG1 and IgG3 on BM nucleated cells increased significantly in BMMNC-Ab IgG positive IRH patients. Disclosures No relevant conflicts of interest to declare.

Influence of the elevated ambient temperature on immunoglobulin G and immunoglobulin G subclasses in sera of Wistar rats

The aim of our research was to examine changes in the immune system of the rats influenced by the elevated ambient temperature. Male Wistar rats were divided, into 2 groups and housed at 20 ? 2?C (n=64, control group) and 35 ? 1?C (n=74, experimental group), during precise timing of 1, 4, 7, 14, 21, and 30 days. All the animals were given food and water ad libitum, and were lighted during 12 hours per day. We have measured IgG, IgG1, IgG2a, IgG2b and IgG2c. The obtained results showed significant elevation in the level of IgG after 4 and 7 days (+32%), IgG2a after 7th (+88%), 14th and 21nd day (+110%), IgG2b after 14 days (+60%) at 35 ? 1?C compared with the control group at 20 ? 2?C. IgG1 level was not affected and IgG2c showed significant decrease after 21st day at 35 ? 1?C. In conclusion, during the elevated ambient temperature the immune system is activated as one of the regulation mechanisms in homeostasis and survival of the population.