scholarly journals PREPARATION AND ANTIGENICITY OF M PROTEIN RELEASED FROM GROUP A, TYPE 1 STREPTOCOCCAL CELL WALLS BY PHAGE-ASSOCIATED LYSIN

1960 ◽  
Vol 112 (1) ◽  
pp. 77-96 ◽  
Author(s):  
Fred S. Kantor ◽  
Roger M. Cole
Keyword(s):  
Cell Walls ◽  
Serum Antibody ◽  
Intradermal Injection ◽  
Exchange Chromatography ◽  
M Protein ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Bactericidal Antibody ◽  
Group A

The lysis of Group A type 1 streptococcal cell walls by phage-associated lysin has been described. In the preparation of lysin, a new semisynthetic non-protein media to support growth of the propagating strain of Group C streptococci was employed. Following lysis of the cell walls, the resulting digest was partially purified by ion-exchange chromatography and ammonium sulfate precipitation. In addition to M protein, the resulting preparation (called lysin M protein) contained the group-specific carbohydrate and the T protein—but did not contain antigens, detectable by precipitin tests, which cross-reacted with absorbed heterologous group or type antisera. Capillary precipitin reactions between the lysin M protein and type-specific antiserum did not occur in the presence of high ionic strength buffers; these buffers did not similarly affect precipitin reactions of acid M protein. Type 1 lysin M protein is shown to be a good antigen. A total of 1.5 mg. injected intradermally in saline produced bactericidal antibody in eight of nine rabbits; when injected in adjuvant in one rabbit, protective serum antibodies developed. Streptococci grown in sera from seven of nine rabbits immunized with lysin M protein demonstrated significantly longer chains than when grown in normal rabbit serum. Antibody as demonstrated by each of these three tests was shown to be type-specific. No local or systemic toxicity was noted following intradermal injection in rabbits of lysin M protein.

scholarly journals STUDIES OF L FORMS AND PROTOPLASTS OF GROUP A STREPTOCOCCI

1959 ◽  
Vol 110 (6) ◽  
pp. 853-874 ◽  
Author(s):  
Earl H. Freimer ◽  
Richard M. Krause ◽  
Maclyn McCarty
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
M Protein ◽  
Group A Streptococci ◽  
Group A ◽  
Isolated Cell ◽  
Streptococcal Strain

L forms of Group A streptococci have been isolated by the use of penicillin gradient agar plates. Osmotically fragile protoplasts of Group A streptococci have been obtained by the use of Group C phage-associated lysin which lyses Group A streptococci and their isolated cell walls. Membranes surrounding these enzymatically derived protoplasts have been isolated, and chemical and immunological studies indicate that they are free of cell wall carbohydrate and M protein. The streptococcal protoplasts reproduce as colonies which are morphologically indistinguishable from streptococcal L forms. Evidence is presented to show that these two streptococcal derivatives are serologically and physiologically related to each other as well as to the parent streptococcal strain from which they were isolated.

scholarly journals Intranasal Immunization with Multivalent Group A Streptococcal Vaccines Protects Mice against Intranasal Challenge Infections

2004 ◽  
Vol 72 (5) ◽  
pp. 2507-2512 ◽  
Author(s):  
Mary A. Hall ◽  
Steven D. Stroop ◽  
Mary C. Hu ◽  
Michael A. Walls ◽  
Mark A. Reddish ◽  
...  
Keyword(s):  
Shigella Flexneri ◽  
Outer Membrane Proteins ◽  
Serum Antibody ◽  
M Protein ◽  
Cholera Toxin B Subunit ◽  
Group A Streptococci ◽  
Cervical Lymph Nodes ◽  
Intranasal Immunization ◽  
Hexavalent Vaccine ◽  
Group A

ABSTRACT We have previously shown that a hexavalent group A streptococcal M protein-based vaccine evoked bactericidal antibodies after intramuscular injection. In the present study, we show that the hexavalent vaccine formulated with several different mucosal adjuvants and delivered intranasally induced serum and salivary antibodies that protected mice from intranasal challenge infections with virulent group A streptococci. The hexavalent vaccine was formulated with liposomes with or without monophosphorylated lipid A (MPL), cholera toxin B subunit with or without holotoxin, or proteosomes from Neisseria meningitidis outer membrane proteins complexed with lipopolysaccharide from Shigella flexneri. Intranasal immunization with the hexavalent vaccine mixed with these adjuvants resulted in significant levels of antibodies in serum 2 weeks after the final dose. Mean serum antibody titers were equivalent in all groups of mice except those that were immunized with hexavalent protein plus liposomes without MPL, which were significantly lower. Salivary antibodies were also detected in mice that received the vaccine formulated with the four strongest adjuvants. T-cell proliferative assays and cytokine assays using lymphocytes from cervical lymph nodes and spleens from mice immunized with the hexavalent vaccine formulated with proteosomes indicated the presence of hexavalent protein-specific T cells and a Th1-weighted mixed Th1-Th2 cytokine profile. Intranasal immunization with adjuvanted formulations of the hexavalent vaccine resulted in significant levels of protection (80 to 100%) following intranasal challenge infections with type 24 group A streptococci. Our results indicate that intranasal delivery of adjuvanted multivalent M protein vaccines induces protective antibody responses and may provide an alternative to parenteral vaccine formulations.

scholarly journals STUDIES ON STREPTOCOCCUS PYOGENES

1959 ◽  
Vol 109 (6) ◽  
pp. 589-600 ◽  
Author(s):  
Hutton D. Slade ◽  
Yoshitami Kimura
Keyword(s):  
Rabbit Antiserum ◽  
M Protein ◽  
Rabbit Serum ◽  
Chemical Fractionation ◽  
Group A Streptococci ◽  
Heterologous Antiserum ◽  
Group A ◽  
Normal Rat ◽  
Group B

Heat-killed cells of Group A streptococci caused death of the adrenalectomized rat. While the adrenalectomized rat readily succumbed to intraperitoneal infection with living cells, death was due primarily to toxicity. The normal rat was highly resistant under either condition. For studies on the toxic materials, the cells of numerous serological types of group A streptococci, and of a Group B and a Group D streptococcus, were extracted with 0.1 N HCl at 100°C. or by sonic oscillation. The extracts, containing macromolecular components, were subjected to chemical fractionation and purification. C substance and M protein of Group A streptococci released from the cell by sonic oscillation were toxic to the adrenalectomized rat in quantities of 1 mg./100 gm. rat. Death usually occurred within 2 hours. On the other hand, C substance and M protein released from the cell with HCl at 100°C. were relatively non-toxic to the adrenalectomized rat. The sonic-extracted C substance of streptococcal Groups B, C, and D was also toxic. The toxic property of the C and M preparations was neutralized in vitro in each case by group and type-specific rabbit antiserum. Heterologous antiserum was without effect. Adrenalectomized rats which received homologous antiserum 18 hours before challenge were also resistant to the toxicity of the C and M preparations. Trypsin destroyed the toxic effect of the M protein preparations and was without effect on the toxicity of the C substance. The R antigen and a nucleoprotein component of Group A streptococci, preparations of protein from Groups B and D streptococci, and coagulase from Staphylococcus aureus were all found to be essentially non-toxicic for the adrenalectomized rat. Large quantities of peptone, crystalline albumin, and rabbit serum were also without effect.

scholarly journals Newly recognized distinctive M protein associated with certain M type 1 group A streptococci.

1975 ◽  
Vol 11 (3) ◽  
pp. 551-555 ◽  
Author(s):  
A E Kholy ◽  
N I Guirguis ◽  
L W Wannamaker ◽  
R M Krause
Keyword(s):  
M Protein ◽  
Group A Streptococci ◽  
Group A

scholarly journals PERSISTENCE OF GROUP A STREPTOCOCCAL CELL WALLS RELATED TO CHRONIC INFLAMMATION OF RABBIT DERMAL CONNECTIVE TISSUE

1967 ◽  
Vol 125 (6) ◽  
pp. 1137-1148 ◽  
Author(s):  
Sarkis H. Ohanian ◽  
John H. Schwab
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Intradermal Injection ◽  
Tissue Response ◽  
Acid Dissociation ◽  
Wall Material ◽  
Phagocytic Cells ◽  
Intact Group ◽  
Polysaccharide Antigens ◽  
Group A

Antibodies specific for the mucopeptide and group-specific C polysaccharide antigens of Group A streptococcal cell walls were prepared by acid dissociation of immune precipitates, and labeled with either fluorescein or 125I. Employing both fluorescent and radioautographic procedures the persistence of the antigens was followed in skin sites injected with cell wall fragments. Both antigens persisted within macrophages for at least 54 days in those animals which developed no chronic tissue response. In animals which did develop chronic nodular lesions the concentration of antigen decreased as the inflammatory process subsided. Lesion activity was thus associated with the presence of cell wall material. The fate of these antigens was also determined following the intradermal injection of intact Group A streptococcal cells. Cell wall antigens persisted in the tissue site considerably longer than morphologically identifiable streptococci, indicating that cell wall fragments are released during dismantling of streptococci in phagocytic cells.

scholarly journals DEMONSTRATION BY IMMUNOFLUORESCENCE OF THE FIXATION OF PERFUSED ANTIBODY TO HUMAN COLLAGEN IN HUMAN KIDNEY

1967 ◽  
Vol 125 (4) ◽  
pp. 595-606 ◽  
Author(s):  
Sidney Rothbard ◽  
Robert F. Watson
Keyword(s):  
Serum Antibody ◽  
Human Kidney ◽  
Basement Membranes ◽  
Human Infant ◽  
Laboratory Animals ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Organs ◽  
Human Collagen ◽  
Testicular Hyaluronidase

Rabbit serum containing antibody to human collagen, perfused through human infant kidneys obtained at autopsy, gives an immunofluorescent reaction with an antigen in the basement membranes of the glomeruli and the tubules. This reaction was shown to be specific by the absence of reaction with normal rabbit serum, antibody to carp collagen, or anti-human collagen serum absorbed with human collagen. Slight cross-reactions were found in the human kidneys with antibody to chicken or rat collagen. There was no evidence that a collagen-like protein in human serum or an antigen common to human erythrocytes and renal glomeruli enters into this immunofluorescent reaction. Perfusion of the kidneys with purified collagenase before the introduction of the antibody to human collagen altered the antigen so that antibody could not be demonstrated in the basement membranes by immunofluorescence. Testicular hyaluronidase used in the same way did not affect the immunofluorescent reaction. This method of perfusion provides another means for studying antigens in human organs by immunofluorescence. These observations in human kidneys extend our earlier findings in laboratory animals and indicate that an antigen, collagen, is also present in human renal glomerular and tubular basement membranes.

Interaction of Group A Type 1 Streptococcal M Protein With Fibrinogen

2009 ◽  
Vol 93B (1-6) ◽  
pp. 201-209
Author(s):  
O. Kühnemund ◽  
J. Havliček ◽  
H. Knöll ◽  
J. Sjöquist ◽  
W. Köhler
Keyword(s):  
M Protein ◽  
Streptococcal M Protein ◽  
Group A

scholarly journals Enzyme-linked immunosorbent assay for streptococcal M protein antibodies

1976 ◽  
Vol 3 (5) ◽  
pp. 501-505
Author(s):  
H Russell ◽  
R R Facklam ◽  
L R Edwards
Keyword(s):  
Immunosorbent Assay ◽  
Cell Walls ◽  
M Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Preliminary Results ◽  
Streptococcal M Protein ◽  
Highly Sensitive ◽  
Bactericidal Antibody ◽  
Human Sera ◽  
M 12

The enzyme-linked immunosorbent assay (ELISA) described by Engvall and Perlmann, which uses antigen-coated tubes and enzyme-labeled anti-immunoglobulin, has been used for the detection of antibodies against streptococcal M protein. The antigen used in the assay was obtained by guanidine extraction of type M-12 streptococcal cell walls followed by hydroxyapatite chromatography. This antigen has the capacity to elicit bactericidal antibodies in rabbits. The results show that the ELISA is specific and highly sensitive for the detection of antibodies in rabbit and human antisera. Preliminary results suggest that, when M-12 antigen is used, the antibodies detected by ELISA are the same antibodies detected in the bactericidal test. The assay has been performed with human and rabbit sera. There was a 96% agreement between bactericidal and ELISA results with rabbit sera and 97.5% agreement with human sera. All bactericidal antibody-positive sera tested thus far yielded positive ELISA results.

scholarly journals In Vitro Preparation And Testing Of Anti-Salmonella Vaccine Against Abortion In Sheep

2017 ◽  
Vol 74 (1) ◽  
pp. 26
Author(s):  
Flore CHIRILA ◽  
George Cosmin NADAS ◽  
Sorin RAPUNTEAN ◽  
Cosmina Maria BOUARI ◽  
Septimiu TOADER ◽  
...  
Keyword(s):  
Cell Walls ◽  
Culture Supernatant ◽  
Serum Antibody ◽  
Brain Heart Infusion ◽  
Broth Culture ◽  
Millipore Filter ◽  
Serum Titer ◽  
Sterility Control

Abstract: In March 2016, the microbiology laboratory of the Faculty of Veterinary Medicine in Cluj-Napoca, 3 type of antisalmonella vaccine for sheep were prepared. For type 1, 24 hours salmonella on brain heart infusion (BHI) broth culture, heat inactivated for 1 hour at 60°C, then with formaldehyde in a concentration of 3 ‰. Variant 2 - the culture supernatant obtained on solid BHI medium, washed with PBS, frozen-thawed 6 times, centrifuged at 4000 rpm for 15 minutes, filtered through 0.2 μm Millipore filter Orange Scientific. Variant 3 - suspension of cell walls remaining after centrifugation suspended in PBS and inactivated with formaldehyde.After the bacterial and fungal sterility control, the three types of vaccine were administered in rabbits by the subcutaneous route at a dose of 1 ml/individual along with 1 ml of plant extract adjuvant (decoction of Ewernia prunastri) sterilized by filtration.There have been two booster inoculations of the initial administration, 7 and 14 days after. Before each dose of vaccine, blood was sampled from the marginal auricular vein in order to control the immunogenicity by anti-somatic ˮOˮ serum antibody (Ab) titration using slow microplate agglutination test (Widal reaction). After three inoculations with the vaccine variant 1, Ab serum titer reached 1/128, and in types 2 and 3, 1/512 after 2 inoculations, decreasing to 1/256 after the second booster administered with no immunomodulator.

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