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001 ocm20346626;

003 OCoLC;

005 20260127093927.0;

008 890915s1987 xx a bm 000 0 eng d;

035 a| (Sirsi) o20346626;

035 a| (OCoLC)20346626;

049 a| EREE;

050 4 a| RC182.S12b| K46x 1987;

100 1 a| Kendall, James E.,c| Jr.,e| author.?| UNAUTHORIZED;

245 14 a| The recognition of cross-reacting, non-flagellar epitopes from Salmonella spp. and Pasteurella spp. using the monoclonal antibody, M467, and polyvalent antisera /c| by James E. Kendall, Jr.;

264 0 c| 1987.;

300 a| [vii], 69 leaves :b| 18 illustrations ;c| 28 cm;

336 a| textb| txt2| rdacontent;

337 a| unmediatedb| n2| rdamedia;

338 a| volumeb| nc2| rdacarrier;

502 b| M.S.c| East Carolina Universityd| 1987;

500 a| Submitted to the faculty of the Department of Biology in partial fulfillment of the requirements of the degree Master of Science.;

500 a| Advisor: A. Mason Smith;

520 3 a| The protein M467 produced by the mouse plasmacytoma MOPC 467 is an IgA class antibody that was shown in previous studies to bind Salmonella spp. flagellin, the monomeric protein of flagella. Further studies have demonstrated that M467 will precipitate antigens from heat extracts of Pasteurella pneumotropica indicating the presence of a common epitope shared by Salmonella flagellin and membrane components present in Pasteurella heat extract. In the present study we have attempted to isolate and characterize the proteins carrying the epitopes present in SaImonella typhimurium and Pasteurella spp. using the M467 protein in double diffusion in agarose gels and various immunoblotting techniques. Bacterial protein patterns were evaluated by SDS-PAGE after Coomassie staining. Isogenic strains of Salmonella typhimurium. ST 25 (flagellated) and ST 26 (non flagellated) were compared to determine if the epitope resided within the membrane of Salmonella. Extracts of Pasteurella multocida P. haemolytica, and P. gallinarium were also tested for activity with M467- During the course of this study it was determined that phenol extracts from the bacteria being investigated contained more available protein than did the heat extracts initially used. Phenol extracts were therefore used in immunoblotting and immunization preparations. The results from immunoblottlng provides evidence that in each of the bacteria shown to be bound by M467, there exists multiple molecular weight proteins that possess the epitope recognized by M467. It is suggested that the epitope present in these gram negative bacterial proteins is a highly conserved peptide sequence that for selective reasons has been retained in the genome of these bacteria throughout their evolution from a common ancestor. It is believed that these bacteria which share a common epitope elicit cross-reacting antibodies that are able to recognize the epitopes of one another. To determine their immunogenic potential, the phenol extracts were used to immunize BALB/c mice. ST 26 and P. multocida phenol extracts were injected i.p. and i.m. using Freund's adjuvant. ELISA assays were performed to detect the production of polyvalent antibodies to the immunogens. Results from the ELISA indicate that the phenol extracts did induce antibody formation and that cross-reactivty was measured in reciprocal assays; ST 26 antisera reacted specifically with ST 26 phenol proteins and with the antigens from P. multocida phenol extract. P. multocida antisera reacted with P. multocida phenol extract and showed cross-reactivity with ST 26 phenol extract. Nonnal mouse serum was used as a negative control. Western blots were performed on the ST 26 and P. multocida phenol extracts using the anti-ST 26 serum and anti-P. multocida serum. Results of these two blots further substantiates that there was cross reactivity between the two phenol extracts of ST 26 and P. multocida. Cross-reacting bands appeared to be identical or very closely related by molecular weight estimation as well as antigenic properties.;

504 a| Includes bibliographical references (leaves 66-69).;

650 0 a| Salmonella.=| ^A44895;

650 0 a| Bacterial diseases.=| ^A6074;

650 0 a| Gram-negative bacteria.=| ^A651109;

650 2 a| Salmonella.0| (DNLM)D012475=| ^A1203944;

650 2 a| Bacterial Infections.0| (DNLM)D001424=| ^A919153;

655 2 a| Academic Dissertation.0| (DNLM)D019478?| UNAUTHORIZED;

650 7 a| Bacterial diseases.2| fast0| (OCoLC)fst00825196;

650 7 a| Gram-negative bacteria.2| fast0| (OCoLC)fst00946119;

650 7 a| Salmonella.2| fast0| (OCoLC)fst01104144;

655 7 a| dissertations.2| aat0| (CStmoGRI)aatgf300028029;

655 7 a| Academic theses.2| fast0| (OCoLC)fst01726453;

655 7 a| Academic theses.2| lcgft;

655 7 a| Thèses et écrits académiques.2| rvmgf0| (CaQQLa)RVMGF-000001173;

700 1 a| Smith, A. Mason,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| East Carolina University.b| Department of Biology.=| ^A637467;

856 41 z| Access via ScholarShipu| http://hdl.handle.net/10342/11217;

949 a| Click on web addressw| asish| joyner101;

949 a| Click on web addressw| asish| hsl111;

994 a| C0b| ERE;

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999 a| MICROFILMw| JMFc| 1i| 223656*l| JMFOTm| JOYNERr| Ys| Yt| JMFOTFIu| 1/22/1990x| MICROFORMz| JMICROFORMo| .STAFF. 0;

999 a| ASK AT SPECIAL COLLECTIONS DESKw| ASISc| 1i| 487796d| 6/7/2001l| JSCJTHDm| JOYNERr| Ys| Yt| JSCJTHDu| 6/7/2001x| ETDz| JSPECCOLLo| .STAFF. 0;

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The recognition of cross-reacting, non-flagellar epitopes from Salmonella spp. and Pasteurella spp. using the monoclonal antibody, M467, and polyvalent antisera / by James E. Kendall, Jr.

Author/creator	Kendall, James E. author.
Other author	Smith, A. Mason, degree supervisor.
Other author	East Carolina University. Department of Biology.
Format	Theses and dissertations
Production	1987.
Description	[vii], 69 leaves : 18 illustrations ; 28 cm
Supplemental Content	Access via ScholarShip
Subjects	Salmonella. Bacterial diseases. Gram-negative bacteria. Salmonella. Bacterial Infections.

Summary	The protein M467 produced by the mouse plasmacytoma MOPC 467 is an IgA class antibody that was shown in previous studies to bind Salmonella spp. flagellin, the monomeric protein of flagella. Further studies have demonstrated that M467 will precipitate antigens from heat extracts of Pasteurella pneumotropica indicating the presence of a common epitope shared by Salmonella flagellin and membrane components present in Pasteurella heat extract. In the present study we have attempted to isolate and characterize the proteins carrying the epitopes present in SaImonella typhimurium and Pasteurella spp. using the M467 protein in double diffusion in agarose gels and various immunoblotting techniques. Bacterial protein patterns were evaluated by SDS-PAGE after Coomassie staining. Isogenic strains of Salmonella typhimurium. ST 25 (flagellated) and ST 26 (non flagellated) were compared to determine if the epitope resided within the membrane of Salmonella. Extracts of Pasteurella multocida P. haemolytica, and P. gallinarium were also tested for activity with M467- During the course of this study it was determined that phenol extracts from the bacteria being investigated contained more available protein than did the heat extracts initially used. Phenol extracts were therefore used in immunoblotting and immunization preparations. The results from immunoblottlng provides evidence that in each of the bacteria shown to be bound by M467, there exists multiple molecular weight proteins that possess the epitope recognized by M467. It is suggested that the epitope present in these gram negative bacterial proteins is a highly conserved peptide sequence that for selective reasons has been retained in the genome of these bacteria throughout their evolution from a common ancestor. It is believed that these bacteria which share a common epitope elicit cross-reacting antibodies that are able to recognize the epitopes of one another. To determine their immunogenic potential, the phenol extracts were used to immunize BALB/c mice. ST 26 and P. multocida phenol extracts were injected i.p. and i.m. using Freund's adjuvant. ELISA assays were performed to detect the production of polyvalent antibodies to the immunogens. Results from the ELISA indicate that the phenol extracts did induce antibody formation and that cross-reactivty was measured in reciprocal assays; ST 26 antisera reacted specifically with ST 26 phenol proteins and with the antigens from P. multocida phenol extract. P. multocida antisera reacted with P. multocida phenol extract and showed cross-reactivity with ST 26 phenol extract. Nonnal mouse serum was used as a negative control. Western blots were performed on the ST 26 and P. multocida phenol extracts using the anti-ST 26 serum and anti-P. multocida serum. Results of these two blots further substantiates that there was cross reactivity between the two phenol extracts of ST 26 and P. multocida. Cross-reacting bands appeared to be identical or very closely related by molecular weight estimation as well as antigenic properties.
General note	Submitted to the faculty of the Department of Biology in partial fulfillment of the requirements of the degree Master of Science.
General note	Advisor: A. Mason Smith
Dissertation note	M.S. East Carolina University 1987
Bibliography note	Includes bibliographical references (leaves 66-69).
Genre/form	dissertations.
Genre/form	Academic theses.
Genre/form	Academic theses.
Genre/form	Thèses et écrits académiques.

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The recognition of cross-reacting, non-flagellar epitopes from Salmonella spp. and Pasteurella spp. using the monoclonal antibody, M467, and polyvalent antisera / by James E. Kendall, Jr.

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