Mouse monoclonal antibodies (P4-2F, P4-5C) against ABO blood group substances in saliva were produced by immunization with ABO blood group active-glycoprotein after ethanol precipitation from heated saliva. These antibodies bound to saliva, irrespective of the ABO blood group and secretor status. Saliva diluted at least 3.2 x 10(4)-fold could be detected by ELISA using these antibodies. Tissue and species specificity of the antibodies was tested by ELISA and counterimmunoelectrophoresis and showed that the antibodies were specific for human saliva. By immunoblotting of the deglycosylated ABO blood group substances it was evident that the epitopes for the antibodies were localized on the core protein of blood group substances in saliva. These antibodies could be extremely suitable reagents for the identification of saliva in medico-legal examinations. Furthermore, they may be used as capture antibodies in sandwich methods for ABO blood grouping of saliva from mixtures of body fluids.
In this paper methods for ABO blood grouping of saliva from mixed body fluids have been established. Monoclonal antibodies to tissue specific epitopes on blood group substances in saliva were used as solid phase antibodies to catch the blood group substances. ABO blood grouping of saliva could be performed by these methods without interference from other body fluids (eg. semen, vaginal secretion, urine, sweat and serum). At least 16,000 and 3,000 fold dilutions of secretor saliva were sufficient for ABO blood grouping by sandwich ELISA and sandwich absorption-elution test, respectively.
Practical methods were developed for ABO blood grouping of semen from mixed body fluids. An monoclonal antibody (P6-5H) which recognizes a tissue-specific epitope on a seminal ABO blood group substance (alpha 2-seminoglycoprotein) was used as a solid phase antibody for selective capture of the seminal ABO blood group substance. ABO blood group epitopes of secretor and non-secretor semen were detected in dilutions of 8 x 10(3)-3.2 x 10(4) and 8 x 10(3)-fold, respectively, by sandwich ELISA. ABO blood group epitopes were also detected in dilutions up to 4 x 10(3)-fold, irrespective of secretor status, by the sandwich absorption-elution test.
Eighty-five cases of infantile asphyxia were examined in relation to the degree of conjunctivae petechial hemorrhages and histological and immunohistochemical findings of the lungs and the pancreas . In very young cases, even in the strangulation cases , conjunctivae petechial hemorrhages were unremarkable and sometimes absent. The lungs showed remarkable to moderate congestion, while the pancreas showed only slight to moderate edema and cell infiltrations.Many pancreata of cases of accidental and homicidal asphyxia had hyperplasia and nesidioblastosis of islet cells. In adult asphyxia cases, remarkable congestion has been the main finding in the lungs and the pancreas. This study shows many similarities between the findings in homicidal suffocation and in genuinely accidental suffocation, both in inspection and on histological examination . So, we here, stressed on the necessity of legal necropsy for various infantile asphyxia cases, in the speculation of the , cause of death, in order to not only study infantile sudden death cause but also not to mis-diagnose genuine accidental asphyxia cases or homicidal cases using suffocation for Sudden Infant Death Syndrome (SIDS).
A novel mouse monoclonal antibody (P4-5C) has been developed which recognizes the core portion of the protein carrying ABO(H) blood group antigens in human saliva. This proved to be specific for human saliva using immunochemical investigations such as enzyme-linked immunosorbent assay, Ouchterlony method and counter-immunoelectrophoresis. By light and electron microscope studies with immunohistochemical techniques using this human saliva-specific P4-5C as primary antibody, it was shown that P4-5C reacted specifically and exclusively with mucus from the mucous gland cells of human salivary glands. P4-5C reacted neither with the mucous gland cells of other primates (hamadryas baboon, Japanese monkey and Rhesus monkey) and four mammals (dog, cat, rabbit and mouse) nor with other human tissues. The epitope on the core portion of the ABO(H)-carrying protein was defined by P4-5C and could be discriminated from the epitope of ABO(H) blood group antigens using immunoelectronmicroscopy, although these 2 epitopes were localized relatively close to each other. The P4-5C monoclonal antibody can be also used for morphological species identification of tissue specimens from submandibular glands.
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