Differential responses of anopheles stephensi [diptera: culicidae] to skin emanations of a man, a cow, and a guinea pig in the olfactometer

Biting habit of mosquitoes plays an important role in the epidemiology of mosquito-borne diseases. Mosquitoes use a set of elaborate sensory modalities to find their preferred hosts by exploiting cues emanating from a nearby host. It has been suggested that the chemical profile of skin can provide further support for anthropophilic mosquito species to find their suitable hosts. This study aimed at revealing the value of skin emanation for a zoophilic species like Anopheles stephensi as a model. Skin emanations of a man, a cow and a Guinea pig were collected by ethanol soaked cottons. Upwind responses of mosquitoes to 100 and 200 micro L of filtered skin materials were non-competitively explored in a dual-choice olfactometer. L-lactic acid and other chemical content of the skin samples were identified by an enzymatic kit and GC-MS, respectively. Unexpectedly, only human skin emanation was resulted in the statistically significant activation and attraction responses of Anopheles. stephensi in the wind tunnel. L-lactic acid content of this skin sample was 10 and 29 times more than the cow and the Guinea pig, respectively. The possible role of lactic acid and a few other identified compounds have been discussed here. An. stephensi showed higher and more specific upwind responses to human skin emanation in the olfactometer. Undoubtedly, the thorough explanation of this unexpected finding needs further investigation. But, if new data verify this result, then, it may be necessary to reconsider the role of skin emanation and thence the human blood index and vectorial capacity of this zoophilic mosquito

Short term reactogenicity of a triple diphtheria-tetanus-whole cell pertussis vaccine in Iranian infants

Immunization against diphtheria, tetanus and pertussis [DTP] has long been applied in Iran using whole cell vaccine. Despite the role of whole cell DTP [DTwP] vaccine in reduction of mortality as a result of disastrous diseases such as diphtheria, tetanus, and pertussis, serious local and systemic complications have been attributed to these vaccines. This study was performed to determine the complications of DTwP vaccine in infants attending some of the health centers of Tehran in 2006-2007. In this prospective study, 330 infants were injected with DTwP vaccine manufactured by Razi Institute of Iran. All subjects received DTwP vaccine at 2, 4, and 6 months of age following the national vaccination schedule of Iran. Reactogenicity was assessed by the parents for 7 days post-vaccination using diary cards. Of the 279 infants who completed the vaccination study, pain was the most frequent local reaction after the primary vaccination [68.1-75.3%]. The mean diameters of the redness and swelling at first day post-vaccination were 2.81 +/- 6.91 and 2.60 +/- 7.93 mm in the first dose, 2.40 +/- 6.25 and 1.94 +/- 5.74 mm in the second dose and 2.24 +/- 5.66 and 2.16 +/- 6.03 in the third dose, respectively. Fever [axillary temperature > 37.5°C] was the most frequently reported systemic reaction during the primary vaccination [53.8-58.8%]. All systemic reactions observed after each dose were either reduced or completely disappeared during a week. The high incident of complications observed following vaccination with this cellular triple vaccine may be related to the formulation or the bacterial cell fragments used in vaccine production

[Designing of a mixed ELISA method for detection of IgM and IgA rheumatoid factors]

Different isotypes of antibody can be produced by immune system after antigen contact. Detection and measurement of different classes of antibody against the antigen is very important in some cases. The aim of this study is designing of an ELISA method on the basis of inhibition of enzyme activity by using a non-competitive inhibitor. Therefore in this study rheumatoid factor is used as a model for the detection of different other classes of antibodies against the antigen.In this cross sectional analytical study, we measured IgM and IgA rheumatoid factors in sera of 10 patients with rheumatoid arthritis and positive latex test, by mixed and routine ELISA. In mixed ELISA the activity of the first conjugated enzyme was blocked by a non-competitive inhibitor after adding the substrate, then the next conjugated antibody, which was specific for another isotype, was added. By optical density, results was comparisoned with routine ELISA.The obtained results showed that the average optical density is lower when compared with routine ELISA, but the difference is not statistically significant. However these two methods did not show any significant difference in quantifying antibody isotypes. Also there is a positive association between mixed and routine ELISA [r =0.9, p=0.001].Lower optical density in mixed ELISA is probably because of stick hindrance by the first conjugate. So, because there is no significant difference between the results of these two types of ELISA, and also no need to repeat the test for each isotype in this method, it is recommended to use the new method instead of the routine one to save time and reagents

case report of recurrent hydrops fetalis due to maternal alloimmunization with Kell antigen

During pregnancy, irregular blood group antibodies originating either from earlier pregnancies or from blood transfusions may severely affect child health. In this report, a case of maternal alloimmunization to Kell antigen is described.The mother had a history of partial mole and four repeated intrauterine fetal death due to hydrops fetalis.Screening of irregular blood group antibodies revealed that she has anti-Kell with the titer of 1:4096. Also in genetic analysis, a C677T homozygous mutation of MTHFR gene was found, which could potentially enhance destructive effects of anti-Kell antibody. The described case emphasizes the importance of being informed about the presence of irregular blood group antibodies during pregnancy which may cause recurrent hydrops

[Monoclonal antibody production against human spermatozoal surface antigens]

As monoclonal antibodies are potential tools for characterization of soluble or cellular surface antigens, use of these proteins has always been considered in infertility and reproduction research. Therefore, in this study, monoclonal antibodies against human sperm surface antigens were produced. To produce specific clones against human sperm surface antigens, proteins were extracted using solubilization methods. Balb/c mice were immunized intraperitoneally with the proteins using complete Freund's adjuvant in the first injection and incomplete Adjuvant in the following booster injections. Hybridoma cells producing ASA were cloned by limiting dilution. Five stable ASA producing hybridoma clones were achieved and their antibody isotypes were determined by ELISA. All the isotypes were of IgG class. Their cross reactivity with rat and mice spermatozoa was examined but they did not have any cross reactivity. The produced antibodies can be used in further studies to characterize and evaluate each of the antigens present on human sperm surface and determining their role in fertilization

[Optimization of Dendritic cells purification from mouse spleen a recommended method for purification of Dendritic cells from reproductive organs]

Dendritic cells [DC] are the principal antigen-presenting cells [APC] responsible for induction of primary immune responses by T lymphocytes. Although DCs are present in most lymphoid tissues, they occur in very low frequency accounting for 0.5% or less of nucleated cells in peripheral lymphoid organs. In the present study, we report the purification of DCs from mouse spleen with high yield and purity using a three-step purification technique including: collagenase digestion of tissue, selection of low-density cells using Optiprep density gradient medium and plastic adherence. By using techniques outlined above, we obtained 5-7x10[7] DC/spleen with purity >/= of 97%. Such large numbers of purified DCs enables us to further document their different characteristics including morphology, immunophenotype and to evaluation of their role in immune system. Finally, since DCs have been reported to be present in all reproductive organs, we suggest that this protocol be used for isolation and purification of DCs from those organs for further in vitro studies

Unresponsiveness to recombinant hepatitis B vaccine in healthy Iranian neonates: Association with HLA antigens

Hepatitis B is an important infectious disease. Since several years ago, mass vaccination against this viral infection has become as part of routine vaccination schedule of Iran. However, some healthy neonates, children and adults fail to generate a protective antibody response after vaccination. To investigate distribution of HLA class-I and class-II antigens in healthy Iranian neonates vaccinated with recombinant hepatitis B vaccine. HLA-typing was performed on peripheral blood lymphocytes isolated from 25 responder and 23 nonresponder [anti-HBs < 10 IU/L] healthy neonates, using the standard microlymphocytotoxicity method. Anti-HBs antibody was quantitated by sandwich ELISA. The frequency of HLA-DR7 [p<0.01], DQ2 [p<0.02] and DR13 [p<0.05] was significantly higher in the nonresponder neonates compared to the responder group. The DR1 and DQ3 antigens were over-represented [p<0.05] in the responder vaccinees, implying positive association with the anti-HBs antibody response. Statistical analysis revealed increased frequencies of B7-DR7-DR53-DQ2 and DR13-DR52-DQ2 haplotypes in the nonresponder neonates [p<0.05]. Conclusions: We found a significant association between lack of antibody response to recombinant hepatitis B vaccine and expression of certain HLA class- II antigens in healthy Iranian neonates

[Production and characterization of a murine monoclonal antibody recognizing a conformational epitope on hCG]

Human chorionic gonadotropin hormone [hCG] belongs to glycoprotein hormones family. Other members of this family include follicle stimulating hormone [FSH], luteinizing hormone [LH] and thyroid stimulating hormone [TSH]. All these hormones consist of a common alfa and a distinct beta subunit. There is a strong similarity between the members of these hormones. Therefore, detection and quantitative measurment of these hormones require production of monoclonal antibodies specific for non-overlapping epitopes on the beta chain or a conformational epitope specific for each hormone. In this study a murine monoclonal antibody against the hCG dimer molecule was produced by hybridoma technology. The specificity of the antibody was assessed by ELISA and Immunoblotting using a panel of highly purified and recombinant forms of glycoprotein hormones including: native hCG and hLH, recombinant hCG, beta hCG, hCG alpha, beta hCG carboxyl terminal peptide covering amino acid residues 109-145 [beta hCG-CTP], recombinant TSH and native FSH, as well as urine proteins [UP]. It was found that the monoclonal antibody reacted with, dimer recombinant and urine purified hCG and hLH, but not with the reduced form of the hormone, nor with recombinant beta hCG, alpha hCG, TSH, native FSH and UP. Using beta hCG-CTP fragment with different concentrations to monitor inhibition of hormone- monoclonal antibody interactions, no interference was observed. This implies that the epitope recognized by the monoclonal antibody is different from that presented by beta hCG-CTP. These results suggest that the monoclonal antibody recognizes a conformational epitope located at the dimer form of hCG molecule and closely associated with the beta subunit of the hormone

IgG subclass analysis of anti-HBs antibodies produced in response to vaccination with recombinant HBsAg and infection with the wild hepatitis B virus

Viral hepatitis, particularly hepatitis B virus [HBV] infection remains a major public health problem worldwide and different profiles of IgG subclasses of anti-HBs antibody have been reported to be produced in individuals infected with HBV and those vaccinated with HBsAg. To evaluate the difference among various profiles of IgG subclasses produced in response to HBV infection and HBsAg observed in HBV positive patients and those vaccinated with HBsAg. Antibody to hepatitis B surface antigen [HBsAg] was purified by affinity chromatography from sera of 24 normal individuals vaccinated with recombinant HBsAg [rHBsAg] and 18 persons infected with hepatitis B virus [HBV]. The isotype and IgG subclass profiles of the purified anti-HBs antibodies were determined by enzyme-linked immunosorbent assay [ELISA] using plates pre-coated with rHBsAg and isotype specific monoclonal antibodies. IgG2 was the predominant subclass of anti-HBs antibody in both groups of individuals followed by IgG4 and IgG1. IgG3 was hardly detectable in the majority of samples tested. No significant differences were observed between the two subject groups with regards to the levels of IgG subclasses and isotypes, suggesting similar antibody responses to rHBsAg and the wild type HBV

Preparation of monoclonal antibody against the common determinants of HBsAg

Subtype-specific monoclonal antibodies are valuable tools for structural analysis of hepatitis B virus surface antigens and epidemiological investigations. Nineteen hybridoma clones producing monoclonal antibodies [MAb] specific for different epitopes of hepatitis B surface antigens [HBsAg] were established from mice immunized with HBsAg of either ayw or adw subtype. The immunizing antigens were purified from serum of carrier individuals or extracts of transgenic plants by affinity chromatography using polyclonal anti-HBs antibody. The specificity of MAbs was determined by enzyme-linked immunosorbent assay and immunoblotting using a panel of purified HBsAg of given subtypes. Out of the 19 MAbs, 9 were found to be specific for the common "a" determinant, and the remainders were specific for either "d" [n=2], "y" [n=5] or "w" [n=3] subtypic determinants. None of the subtype-specific MAbs displayed cross-reactivity with the other major subtypic epitopes. These MAbs have potential as monospecific reagents for subtyping HBsAg in carrier individuals for epidemiological and experimental investigations

Cytogenetic analysis of malignant B-cells from iranian patients with chronic lymphocytic leukemia

Lymphocytes of 37 Iranian patients with B-cell chronic lymphocytic leukemia were stimulated with different mitogens i.e., phorbol myristate acetate [PMA], pokweed mitogen and Staphylococcus aureus Cowan I. Adequate metaphases were obtained in 23 subjects following PMA stimulation. Cytogenetic analysis of G-banded mitosis showed both numerical and structural abnormalities. The most frequent abnormality observed in this study involved chromosome 21 [-21 in four cases and +21 in three cases]. Trisomy 12 and 15 as well as structural abnormalities and other alterations related to chromosomes 2, 3, 6, 7, 10, 12, 18 and 19 were also observed. The high frequency of numerical abnormalities involving chromosome 21 found in this study may be considered as a characteristic feature of B-CLL in Iran

Hepatitis C virus infection in patients with lymphoproliferative disorders

Involvement of hepatitis C virus [HCV] in pathogenicity of some lymphoproliferative disorders has been recently proposed. In the present study, the frequency of anti-HCV antibody [Ab] was determined in 42 patients with essential mixed cryoglobulinemia [EMC], 45 with multiple myeloma [MM] and 23 with B-cell chronic lymphocytic leukemia [B-CLL]. Specific antibodies to HCV antigens were detected by enzyme-linked-immunosorbent assay and positive results were confirmed by recombinant immunoblot assay. Our results demonstrated anti-HCV positivity in 69% of EMC, 11% of MM and 4.3% of B-CLL samples tested. Considering the low prevalence of HCV infection in normal population [<1%], these data confirm and extend previous reports on the association of HCV infection with EMC and perhaps with MM

Immunogenicity of recombinant hepatitis B vaccine in Iranian neonates: high frequency of unresponsiveness independeut of the carrier state of mothers

Immunogenicity of a recombinant hepatitis B vaccine was evaluated in a number of Iranian neonates. Of 1176 neonates vaccinated with triple doses of the vaccine at 0, 1 and 6 months, 7.4% failed to respond and did not develop a protective titer of anti-HBs antibody [>/= 10 IU/1]. Titration of anti-HBs antibody allowed classification of both responder and non-responder vaccinees into negative or borderline non-responders and low, intermediate or high responders, respectively. Unresponsiveness to the vaccine was similarly represented in both male and female vaccinees. HBs antigen was negative in all responder and non-responder neonates tested and their mothers, suggesting lack of correlation between unresponsiveness of the neonates to recombinant HBs antigen and the carrier status of the mothers