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Unrelated Antibodies Peptide Mimotope Can Bind to Two Capsular Polysaccharide of 6B Serotype

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Unrelated Antibodies Peptide Mimotope Can Bind to Two Capsular Polysaccharide of 6B Serotype Peptide Mimotopes of Pneumococcal Capsular Polysaccharide of 6B Serotype: A Peptide Mimotope Can Bind to Two Unrelated Antibodies This information is current as of October 1, 2021. Jeon-Soo Shin, Jigui Yu, Jisheng Lin, Linghao Zhong, Kara L. Bren and Moon H. Nahm J Immunol 2002; 168:6273-6278; ; doi: 10.4049/jimmunol.168.12.6273 http://www.jimmunol.org/content/168/12/6273 Downloaded from References This article cites 30 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/168/12/6273.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 1, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Peptide Mimotopes of Pneumococcal Capsular Polysaccharide of 6B Serotype: A Peptide Mimotope Can Bind to Two Unrelated Antibodies1 Jeon-Soo Shin,* Jigui Yu,‡ Jisheng Lin,‡ Linghao Zhong,† Kara L. Bren,† and Moon H. Nahm2‡§ Two groups of bacteriophage clones displaying the antigenic properties of serotype 6B pneumococcal capsular polysaccharide (PS) were obtained from different phage libraries expressing random heptameric peptides. One group, biopanned with a mouse mAb (Hyp6BM1), is comprised of 17 phage clones expressing 10 unique sequences of linear peptides. The other group, selected with another mAb (Hyp6BM8), contained six clones, all of which expressed the identical circular peptide. Phage clones expressing the linear peptides (e.g., PhaM1L3) bound only to Hyp6BM1, but not other 6B PS-specific mAb, and their binding could be inhibited Downloaded from with pneumococcal capsular type 6B PS only. In contrast, a phage clone expressing the circular peptide (PhaM8C1) cross-reacted with several other 6B PS-specific mAbs, and their binding could be inhibited with pneumococcal capsular PS of 6A and 6B serotypes. Two short peptides, PepM1L3 and PepM8C1, reflecting the peptide inserts of the corresponding phage clones, could inhibit the binding of the two clones to their respective mAb. Interestingly, the peptide insert in PhaM8C1 was identical to that in PhaB3C4, a previously reported mimotope of ␣(238) polysialic acid, Neisseria meningitidis group B PS. Indeed, PhaM8C1 bound to HmenB3 (a meningococcal Ab), and their association could be inhibited with ␣(2–8) polysialic acid, but not with 6B PS. http://www.jimmunol.org/ Conversely, ␣(2–8) polysialic acid could not inhibit the binding of PhaM8C1 to Hyp6BM8. The two-dimensional nuclear magnetic resonance studies indicate that PepM8C1 peptide can assume several conformations in solution. The ability of this peptide to assume multiple conformations might account for its ability to mimic more than one Ag type. The Journal of Immunology, 2002, 168: 6273–6278. s different peptides can assume diverse conformations, a should be useful for studying differences in immune mechanisms large peptide library expressing random sequences used by PS and protein Ags. In addition, the peptide mimics should contain peptides that can conformationally should, unlike PS, elicit T cell help, immune memory, and strong A by guest on October 1, 2021 mimic virtually any ligand or Ag. This idea has been shown to Ab responses. Also, the peptides should be easier to manufacture have practical application, with the recent demonstration that pep- and modify than PS Ags (7, 8). Thus, the mimotopes may be useful tide mimics of antigenic specificities can be efficiently identified as vaccine components. However, despite reports of successful using large peptide libraries. For instance, random peptides can be mimotope vaccines (4, 9–11), mimotopes often either are poorly displayed on a coat protein of a bacteriophage, and biopanning can immunogenic, elicit ineffective Abs, or induce B cell memory for be used to isolate phage clones expressing peptides of a desired ineffective Abs. Consequently, there is a need for further studies of binding specificity (1–3). This approach has been successfully peptide mimics of PS Ags before peptide mimic vaccines can used to identify various antigenic mimics (mimotopes). The mi- reach their full potential. motopes have been used to examine Ab fine specificity (4), to Streptococcus pneumoniae is a well-known pathogen, causing examine the requirement for B cell stimulation by Ags (5), and to several serious diseases in young children and the elderly (12). As elicit desirable Abs (6). a result, Ab responses to pneumococcal PS Ags have been exten- Peptides mimicking polysaccharide (PS)3 Ags can be readily sively studied in the past. The peptide mimics can be easily com- produced and used various ways. The peptide mimics of PS Ags pared with other pneumococcal Ags for their immunogenicity and for inducing protective Abs. Also, there is a need for more effec- tive pneumococcal vaccines. The widely available pneumococcal *Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea; vaccine containing capsular PS from 23 common serotypes (13) is †Department of Chemistry, University of Rochester, Rochester, NY 14642; and De- not immunogenic in young children, and its effectiveness is re- partments of ‡Pathology and §Microbiology, School of Medicine, Division of Labo- ratory Medicine, University of Alabama, Birmingham, AL 35294 duced among the elderly (14). A new conjugate vaccine, although Received for publication January 14, 2002. Accepted for publication March 27, 2002. effective in young children (15), is not effective among the elderly (16) and is expensive to manufacture. We have studied the peptide The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance mimics of pneumococcal capsular PS and have identified a peptide with 18 U.S.C. Section 1734 solely to indicate this fact. that can mimic pneumococcal PS as well as meningococcal PS. 1 This work is supported by National Institutes of Health Grant AI-31473 (to M.H.N.) and a Howard Hughes Biomedical Research Support Program grant (to K.L.B.). Materials and Methods 2 Address correspondence and reprint requests to Dr. Moon H. Nahm, Department of Antibodies Pathology, School of Medicine, Division of Laboratory Medicine, University of Al- abama, 845 19th Street South, BBRB 614, Birmingham, AL 35294. E-mail address: HmenB1 and HmenB3 are IgM mAb specific for Neisseria meningitidis [email protected] group B capsular PS (17). The 101.4.1 is a mAb to N-acetyl-␤-D-glu- 3 Abbreviations used in this paper: PS, polysaccharide; 2-D, two-dimensional; NMR, cosamine from M. Cunningham (University of Oklahoma, Oklahoma City, nuclear magnetic resonance. OK) (18). Hyp6A1 is a mouse IgM mAb specific for pneumococcal 6A PS. Copyright © 2002 by The American Association of Immunologists, Inc. 0022-1767/02/$02.00 6274 A PEPTIDE MIMOTOPE BINDING TO TWO DIFFERENT Abs Dob1 is a human IgG2 mAb binding to 6B PS (19). Hyp6BM1, Hyp6BM7, was identified with an underline and derived from the peptide inserts of Hyp6BM8, and Hyp6BM10 are mouse IgM mAb specific for pneumococ- PhaB3C1 (17), PhaM1L3, PhaM1L9, or PhaM8C1. The amino acid Y or C cal 6B PS (20). Dob1, Hyp6BM7, Hyp6BM8, and Hyp6BM10 cross-react at the N-terminal was added for the purpose of radiolabeling or conjugation with 6A PS, an isopolymer of 6B PS. Hyp6BM1 does not cross-react with with carrier protein. The remaining amino acids in either sides of the core 6A PS, and Hyp6A1 does not cross-react with 6B PS (20). Hyp6BM1 and peptide were derived from the sequence of the phage protein pIII flanking Hyp6BM8 were used for biopanning, and the mAb were purified from the the peptide inserts. These peptides were synthesized by Biosynthesis mouse ascites by (NH4)2SO4 precipitation and by chromatography over a (Lewisville, TX) or Biomolecules Midwest (Waterloo, IL). column of Sephacryl S-300HR (Pharmacia Biotech, Uppsala, Sweden). NMR spectroscopy Production of phage clones expressing the mimotopes Solutions (2.5 mM) of NLpeptide6 and NLpeptide9 were prepared in PBS. Two phage libraries from New England Biolabs (Beverly, MA) were used The 1-H TOCSY spectra of both peptides were obtained on a 500-MHz for our study. One contained linear peptides composed of 7 random aa, and Varian INOVA nuclear magnetic resonance (NMR) spectrometer (Varian, the other contained circular peptides of 9 aa; circularization was achieved Palo Alto, CA) using a standard pulse sequence (23, 24). Data were pro- by a covalent bond between the cysteines at positions 1 and 9, and the 7 aa cessed and analyzed using FELIX 97 software (Accelrys, San Diego, CA). between the two cysteines are randomly chosen. Biopanning the phage libraries was performed as described (17). Briefly, 60-mm petri dishes Results (Nunc, Roskilde, Denmark) were coated with mAb at a concentration of Selection of phage clones and analysis of their binding 100 mg/L in 0.1 M NaHCO3 (pH 8.6). The mAb-coated petri dishes were blocked with 0.5% BSA in 0.1 M NaHCO3 and washed with 0.1% TBST properties (50 mM Tris-Cl (pH 7.5), 150 mM NaCl). For each biopanning cycle, ϳ2 ϫ 1011 PFU phage were placed in the petri dish and incubated for 30 Biopanning of the phage library displaying the linear peptides with min.
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