دراسة مناعية لمستضد متعدد السكريد للفطر Aspergillus niger في الارنب == Immunological Study of Aspergillus Niger Polysaccharide Antigen In Rabbit
Author name:
غدير كاظم عبود الخفاجي
Supervisor name:
نداء شهاب حمد | فريال جميل عبد
General topic:
Biology
Specific topic:
Microbiology
Degree:
Master
University:
University of Babylon - College Of Science - Department Of Biology
Language:
Arabic
University location:
Babylon
First pages:
24T2748 - p.pdf
Abstract:
جمعت خلال هذه الدراسة 50 عينة تمثلت ب 25 عينة تربة و25 عينة قشع من اشخاص مصابين بالتدرن الرئوي والوافدين لمركز التدرن والامراض الصدرية في محافظة بابل خلال المدة المحصورة للفترة من تشرين اول 2013 ولغاية شباط 2014 تم خلالها عزل الفطر Aspergillus niger وبنس | fifty specimens were collected, 25 samples were collected from soil while the other 25 sample were collected from sputum of pateints with Tuberculosis, who admitted to thoracic diseases center in Babylon province in period October 2013 to Febreury 2014. Aspergillus niger Has been Isolated from these sample with 56% appearance in soil & 48% in sptum of TB pateints.these isolated were examined micro and macroscopically. purified Fungi was culture on potato dextrose broth, after incubation period polysaccharide antigen of A.niger cell wall was prepared for isolated from soil and TB patients suptum. Molish, protein and glucose test to provide that these antigen are polysaccharide, then immunuization of local rabbit of Orcyctalagus conninculus age 3 - 5 months & 1 - 1.5 Kg for males only (3 animals for each antigen) After the immunization period (one month) some of local and systemic immune response parameters (cellular and humoral) to test animals were studied and compared with control animals which immunized with normal saline. The results of study showed that the polysaccharide antigen isolated from these two strains lead to stimulate systemic humaral immune response which had been studied by using direct agglutination test for appendix immunoglobulin passive haemagglutination test (both were used to determined antibody titer), radialimmunodiffusion test in order to determine level of concentration of immunoglobulins (IgG, IgM) and complement part (C3, C4). where The results of direct agglutination titer of animals immunized with polysaccharide antigen 128 and 1 for control animals and heamagglutination tests titer of animals immunized with polysaccharide antigen serum (2560). These results were higher compared with control group titer (10) and this is also true for immunoglobulin and concentration of IgG, IgM for immunized animals compared to control one. In addition to that The concentration of C3 and C4 had been increased in immunized animals compared to the control. cellular immunological response had been measured by using rabbit Neutrophil Elastase Kit. The eresult show an increase in the level of phagocytosis in animals immunoized with polysaccharide antigen isolated from A.niger cell wall of TB pateints 0.37±0.03 pg/dl while decreased in animals immunoized with polysaccharide of soil A.niger 0.72294±0.25548pg/dl compared with control animals 0.6±0.16386 pg/dl. on the other hand the humaral adaptive immune response had been identified by measuring cytokine IL - 2, IL - 10 in serum rabbit, the level of IL - 2 in rabbit immunoized with polysaccharide antigen soure TB patients and soil is 5.4129±1.08979 and 4.2641±0.62997 pg/dl respectively compared with control animals 3.9327±0.55587 pg/dl. The level of IL - 10 had been dropped for immunized animals with A.niger of TB pateints and from soil 4.8762±1.15104 and 8.2632±4.31933 pg/dl while the control animals having the highest level 10.2576±5.23356 pg/dl among the groups. IgE concentration measured significant difference at p>0.05 in immunized animals of A.niger in both TB pateints and soil 1.1701±0.05811 and 1.0770±0.19355 ml/Iu compared with control where IgE0.8723±0.0849ml/Iu. Miotic index of bone morrow immunized and control animals show mean difference is signification at p>0.05.where the result demonstrate that the polysaccharide antigen fissed the bone marro cells compared to controlled one. In order to identify the viability of antigen in protection. challenge dose test made by injection animals with 108 spore/ml of A.niger conidia which prove their protection in immunized animals while the control animals undergo many morphological and behavioral changes after 5 - 7 days of their injection.I addition that the size of lung and spleen increased when compared with immunized one. The histological culture of lung and spleen of challenge test animals in order to identify the ability and speed of their body to evadicate and prevent infection, display that A.niger coloni appeared for control animals (6 rabbit) while disappeared in culture of immunized one. Skin test also performed to experimental animals which indicate that polysaccharide antigen of A.niger interact with hypersensitivity test of rabbit, where the level of late hypersenetivity increased clearly with propability of p>0.05 with the highest level after 24 hours of beginning test compared to controlled one.
