共查询到20条相似文献,搜索用时 31 毫秒
1.
Yoav I. Henis Orit Gutman 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1983,762(2):281-288
We have studied the diffusion and aggregation of H-2Kk antigens labeled with a fluorescent anti-H-2Kk monoclonal antibody (IgG) on mouse splenic lymphocytes, employing fluorescence photobleaching recovery and fluorescence microscopy. The H-2Kk antigens were initially distributed homogeneously on all lymphocytes. Upon antibody binding, sub-micron patches were formed on 50–60% of the cells. A lateral diffusion coefficient, D, of 7.1·10?10 cm2/s and a mobile fraction of 0.73 were found for H-2Kk antigens on diffusely-labeled cells, while these antigens were immobile (D?5·10?12 cm2/s) on patched cells. The patched and nonpatched sub-populations did not correspond to B- and T-lymphocytes. Subjection to low temperature or treatment with NaN3 or cytoskeleton-disrupting drugs did not affect the diffusion or patching of H-2Kk, indicating no involvement of metabolic energy or drug-sensitive cytoskeletal components. These findings could be related to the interactions of H-2 antigens on the cell surface, and to the different susceptibilities of various cells to lysis by cytotoxic T-cells. 相似文献
2.
Zhenping Zhu Jaroslav Kralovec Tarunendu Ghose Molly Mammen 《Cancer immunology, immunotherapy : CII》1995,40(4):257-267
The anthracyclin antineoplastic agent doxorubicin (Adriamycin) was linked by four different methods of linkage to DalB02, an IgG1 murine monoclonal antibody (mAb) against surface-associated antigens on human chronic lymphocytic leukaemia (CLL) B cells. All the four conjugates fully retained the immunoreactivity of the parent DalB02. When the inhibitory effect of these conjugates was evaluated in vitro against the target D10–1 cells (a clone derived from an Epstein-Barr-virus-transformed human CLL B cell line that binds DalB02) it was observed that one conjugate was more potent than the free drug but the others were not. When131I-labelled unmodified DalB02 and the131I-labelled DalB02-containing conjugate that was found to be potent were injected i.v. into nude mice bearing a subcutaneous D10–1 xenograft, the percentages of the injected dose (%ID) of both131I-DalB02 and the131I-DalB02-containing conjugate that localized in the tumour were much higher than the %ID of the respective preparations that localized in normal tissues of D10–1-xenografted mice. The systemic toxicity of the conjugate was less than that of the free drug. At an equitoxic dose level, this conjugate was a more effective inhibitor of established D10–1 xenografts than the free drug.This study was supported by grants from the Medical Research Council of Canada (grant MT 10964) and the Cancer Research Society Inc., Montreal, Canada 相似文献
3.
Asanuma H Matsumoto-Takasaki A Suzuki Y Tamura S Sata T Kusada Y Matsushita M Fujita-Yamaguchi Y 《Biochemical and biophysical research communications》2008,366(2):445-449
Anti-influenza hemagglutinin (HA) Fabs were isolated from a phage display library using purified HA of influenza virus A/Puerto Rico/8/34 (PR8; H1N1) as an antigen. Four Fab clones displaying a 25-50-fold higher binding signal to PR8 HA than the control were selected for further analysis and comparison with anti-PR8 monoclonal antibody (mAb). All four Fabs and mAb recognized the PR8 HA under non-reducing conditions but rarely bound to reduced PR8 HA. Inhibition of influenza virus infection on MDCK cells was observed with Fab1 and mAb in a dose-dependent manner while Fab3 and 4 exhibited only a partial inhibitory effect. Moreover, Fab1 clone and mAb exhibited cross-reactivity with the A/Peking/262/95 (A/Peking; H1N1) strain. The inhibitory effects of mAb on both influenza strains were more potent than Fab1, which is likely attributed to its higher affinity for the antigen. SPR analyses, in fact, revealed that Fab1 and mAb have KD of 1.5 × 10−8 and 3.2 × 10−9 M, respectively. These results strongly suggest that phage library-derived Fabs can be readily prepared and that such HA-specific Fabs with inhibitory action on influenza infection may be used to treat influenza patients. 相似文献
4.
Xuchu Que George F. Widhopf II Shahzada Amir Karsten Hartvigsen Lotte F. Hansen Douglas Woelkers Sotirios Tsimikas Christoph J. Binder Thomas J. Kipps Joseph L. Witztum 《PloS one》2013,8(6)
The immunoglobulins expressed by chronic lymphocytic leukemia (CLL) B cells are highly restricted, suggesting they are selected for binding either self or foreign antigen. Of the immunoglobulin heavy-chain variable (IGHV) genes expressed in CLL, IGHV1-69 is the most common, and often is expressed with little or no somatic mutation, and restricted IGHD and IGHJ gene usage. We found that antibodies encoded by one particular IGHV1-69 subset, designated CLL69C, with the HCDR3 encoded by the IGHD3-3 gene in reading frame 2 and IGHJ6, specifically bound to oxidation-specific epitopes (OSE), which are products of enhanced lipid peroxidation and a major target of innate natural antibodies. Specifically, CLL69C bound immunodominant OSE adducts termed MAA (malondialdehyde–acetaldehyde-adducts), which are found on apoptotic cells, inflammatory tissues, and atherosclerotic lesions. It also reacted specifically with MAA-specific peptide mimotopes. Light chain shuffling indicated that non-stochastically paired L chain of IGLV3-9 contributes to the antigen binding of CLL69C. A nearly identical CLL69C Ig heavy chain was identified from an MAA-enriched umbilical cord phage displayed Fab library, and a derived Fab with the same HCDR3 rearrangement displayed identical MAA-binding properties. These data support the concept that OSE (MAA-epitopes), which are ubiquitous products of inflammation, may play a role in clonal selection and expansion of CLL B cells. 相似文献
5.
Andrey S. Dobroff Elaine G. Rodrigues Maria A. Juliano Dayson M. Friaça Ernesto S. Nakayasu Igor C. Almeida Renato A. Mortara Jacqueline F. Jacysyn Gustavo P. Amarante-Mendes Walter Magliani Stefania Conti Luciano Polonelli Luiz R. Travassos 《Translational oncology》2010,3(4):204-217
Malignant melanoma has increased incidence worldwide and causes most skin cancer-related deaths. A few cell surface antigens that can be targets of antitumor immunotherapy have been characterized in melanoma. This is an expanding field because of the ineffectiveness of conventional cancer therapy for the metastatic form of melanoma. In the present work, antimelanoma monoclonal antibodies (mAbs) were raised against B16F10 cells (subclone Nex4, grown in murine serum), with novel specificities and antitumor effects in vitro and in vivo. MAb A4 (IgG2ak) recognizes a surface antigen on B16F10-Nex2 cells identified as protocadherin β13. It is cytotoxic in vitro and in vivo to B16F10-Nex2 cells as well as in vitro to human melanoma cell lines. MAb A4M (IgM) strongly reacted with nuclei of permeabilized murine tumor cells, recognizing histone 1. Although it is not cytotoxic in vitro, similarly with mAb A4, mAb A4M significantly reduced the number of lung nodules in mice challenged intravenously with B16F10-Nex2 cells. The VH CDR3 peptide from mAb A4 and VL CDR1 and CDR2 from mAb A4M showed significant cytotoxic activities in vitro, leading tumor cells to apoptosis. A cyclic peptide representing A4 CDR H3 competed with mAb A4 for binding to melanoma cells. MAb A4M CDRs L1 and L2 in addition to the antitumor effect also inhibited angiogenesis of human umbilical vein endothelial cells in vitro. As shown in the present work, mAbs A4 and A4M and selected CDR peptides are strong candidates to be developed as drugs for antitumor therapy for invasive melanoma. 相似文献
6.
Ana-Carolina Martinez-Torres Claire Quiney Tarik Attout Helo?se Boullet Linda Herbi Laura Vela Sandrine Barbier Danielle Chateau Elise Chapiro Florence Nguyen-Khac Frédéric Davi Magali Le Garff-Tavernier Roba Moumné Marika Sarfati Philippe Karoyan Hélène Merle-Béral Pierre Launay Santos A. Susin 《PLoS medicine》2015,12(3)
7.
Alexander Knuth Wolfgang Dippold Karl-Hermann Meyer zum Bueschenfelde 《Cellular immunology》1984,83(2):398-403
Cytotoxic T lymphocytes (CTL) for autologous malignant melanoma in culture of a patient AV were induced by restimulation of PBL (peripheral blood leukocytes) with AV melanoma cells in vitro and subcultured in interleukin 2 (IL-2) conditioned media. Monoclonal antibodies detecting six antigenic systems on melanoma cell surfaces were tested for blocking activity on the effector function of subcultured cytolytic T lymphocytes for autologous melanoma cells. The monoclonal antibodies R24 (γ3), specific for the GD3 disialoganglioside on melanoma cell surfaces and I24 (γM), detecting a similar antigenic determinant, blocked autologous T lymphocytotoxicity for malignant melanoma cells on the target level. The effector function of alloantigen activated cytolytic T lymphocytes generated by coculture of allogeneic PBL with Epstein-Barr virus (EBV) transformed AV B lymphocytes, was blocked by monoclonal antibody R24 when tested against AV melanoma targets, but not when tested against AV B lymphocyte targets. It is concluded that blocking by mAb R24 occurs in this system as a nonspecific effect, unrelated to the specific target antigen recognition by cytotoxic T lymphocytes. Steric hindrance or antibody induced membrane changes may account for the blocking effect of monoclonal antibody R24. 相似文献
8.
Lymphocytes from rodents cultured on syngeneic fibroblasts become cytotoxic against syngeneic but not against allogeneic target cells. We investigated whether known antigens are involved in the phenomenon and the data indicate that H-2 antigens must be shared between sensitizing fibroblasts and responder lymphocytes to generate autocytotoxic cells. Furthermore, the cytotoxicity of autosensitized lymphocytes is restricted to target cells identical with respect to theK and/orI regions. F1 hybrid lymphocytes cultured on parental fibroblasts develop cytotoxicity towards sensitizing cells. In contrast, parental lymphocytes cultured on F1 hybrid fibroblasts will not damage the F1 cells, although they are cytotoxic against both syngeneic and allogeneic parental cells. In addition, parental or F1 hybrid lymphocytes cultured on parental fibroblasts are not cytotoxic against F1 hybrid target cells. Fibroblasts heterozygous for theK end only, are also resistant to the cytotoxic action of such lymphocytes. Thus it seems that H-2 antigens, specifically theK end, antigens have a significant role in the phenomenon of autosensitization. 相似文献
9.
Hiroyuki Satofuka Satoshi Amano Toshiaki Fukui Haruyuki Atomi Masahiro Takagi Tadayuki Imanaka 《Biotechnology letters》2000,22(17):1423-1428
Phytochelatins (PCs, (Glu-Cys)n-Gly, n = 2–11) are metal ions-binding peptides produced by plant, algae and fungi. Antibodies that recognize PCs were induced by the injection of Balb/c mice with a multiple antigen peptide consisting of PC6(MAP-PC6). One stable hybridoma producing a monoclonal antibody (mAb), designated as 4-9C, was established. The DNA sequences of the heavy and light chain variable regions of the 4-9C mAb were determined. The 4-9C mAb had a smaller equilibrium dissociation constant (K
d) towards Cu-, Zn- and Ni-PC7complexes than those towards other metal-PC7complexes and free PCs. 相似文献
10.
There is a constant need to identify novel inhibitors to combat β-lactamase-mediated antibiotic resistance. In this study, we identify three penicillinase-binding peptides, P1 (DHIHRSYRGEFD), P2 (NIYTTPWGSNWS), and P3 (SHSLPASADLRR), using a phage display library. Surface plasmon resonance (SPR) is utilized for quantitative determination and comparison of the binding specificity of selected peptides to penicillinase. An SPR biosensor functionalized with P3-GGGC (SHSLPASADLRRGGGC) is developed for detection of penicillinase with excellent sensitivity (15.8 RU nM−1) and binding affinity (KD = 0.56 nM). To determine if peptides can be good inhibitors for penicillinase, these peptides are mixed with penicillinase and their inhibition efficiency is determined by measuring the hydrolysis of substrate penicillin G using UV–vis spectrophotometry. Peptide P2 (NIYTTPWGSNWS) is found to be a promising penicillinase inhibitor with a Ki of 9.22 μM and a Ki′ of 33.12 μM, suggesting that the inhibition mechanism is a mixed pattern. This peptide inhibitor (P2) can be used as a lead compound to identify more potent small molecule inhibitors for penicillinase. This study offers a potential approach to both detection of β-lactamases and development of novel inhibitors of β-lactamases. 相似文献
11.
Zhou Qinwei Desai Smruti A. Wang Xinhui Noronha Elvyra J. Neri Mariangela Ferrone Soldano 《International journal of peptide research and therapeutics》1999,6(1):77-86
Summary Utilizing phage display peptide libraries, we have identified and mapped the antigenic determinants recognized by mouse monoclonal
antibodies (mAb) on two sets of immunologically important molecules, HLA class I and class II antigens. Anti-HLA class I mAb
TP25.99 recognizes a conformational and a linear determinant on distinct regions of the HLA class I α3 domain. Anti-HLA class
I mAb HO-4 recognizes a conformational determinant on the α2 domain of HLA-A2 and A28 allospecificities. Anti-HLA-DR1,-DR4,-DR6,-DR8,-DR9
mAb SM/549 recognizes a conformational determinant on the β chain of HLA class II antigens. These results indicate the versatility
of phage display peptide libraries to characterize antigenic determinants recognized by anti-HLA mAb. 相似文献
12.
H C Ertl 《Cellular immunology》1981,62(1):38-49
This paper investigates kinetics and antigenic and genetic requirements for transfer of delayed-type hypersensitivity (DTH) reactions against Sendai virus. Kinetics of sensitization of effector cells are similar to those described in other virus systems. Maximum DTH response is elicited in the footpad 7 days after sensitization; maximal increase in footpad thickness is found approximately 24 hr after injection of virus and transfer of immune lymphocytes. DTH effector cells are Ig? and are susceptible to treatment with anti-theta antibody and complement. After transfer of immune lymphocytes a DTH reaction can be elicited by infectious virus, uv light-inactivated virus or cell-cultured Sendai virus which lacks fusion capacity. Requirements of H-2 compatibility are dependent on the biological nature of the virus preparation used for challenge: High doses of infectious or uv light-inactivated Sendai virus require K, D, or IA region compatibility; low concentrations of infectious virus require K and/or D region compatibility, while cell-cultured fusion-negative Sendai virus requires IA region homology. At the level of induction K, D region-restricted DTH-effector T lymphocytes (Td) and cytolytic T lymphocytes (Tc) are stimulated to a greater extent by infectious virus than by uv light-inactivated virus. Furthermore, stimulation of these T lymphocytes is dependent on the route chosen for immunization: intravenous (iv) injection is superior to intraperitoneal (ip) injection; subcutaneous (sc) injection causes the lowest degree of sensitization. IA region-restricted Td lymphocytes are stimulated to comparable amounts by infectious or uv light-inactivated Sendai virus independent of the route of immunization. Td lymphocytes, which require IA region compatibility and Td lymphocytes which require K or D region compatibility can be distinguished by their Lyt phenotype, e.g., IA region-restricted Td lymphocytes are characterized by Lyt-1+2? antigens, while K or D region-restricted Td lymphocytes are phenotypically Lyt-1(+)2+. 相似文献
13.
Johanna H Kattenberg Inge Versteeg Stephanie J Migchelsen Iveth J González Mark D Perkins Petra F Mens Henk DFH Schallig 《MABS-AUSTIN》2012,4(1):120-126
Currently available rapid diagnostic tests (RDTs) for malaria show large variation in sensitivity and specificity, and there are concerns about their stability under field conditions. To improve current RDTs, monoclonal antibodies (mAbs) for novel malaria antigens have been developed and screened for their possible use in new diagnostic tests. Three antigens, glutamate rich protein (GLURP), dihydrofolate reductase-thymidylate synthase (DHFR-TS) and heme detoxification protein (HDP), were selected based on literature searches. Recombinant antigens were produced and used to immunize mice. Antibody-producing cell lines were subsequently selected and the resulting antibodies were screened for specificity against Plasmodium falciparum and Plasmodium vivax. The most optimal antibody couples were selected based on antibody affinity (expressed as dissociation constants, KD) and detection limit of crude antigen extract from P. falciparum 3D7 culture. The highest affinity antibodies have KD values of 0.10 nM ± 0.014 (D5) and 0.068 ± 0.015 nM (D6) for DHFR-TS mAbs, 0.10 ± 0.022 nM (H16) and 0.21 ± 0.022 nM (H18) for HDP mAbs and
0.11 ± 0.028 nM (G23) and 0.33 ± 0.093 nM (G22) for GLURP mAbs. The newly developed antibodies performed at least as well as commercially available histidine rich protein antibodies (KD of 0.16 ± 0.13 nM for PTL3 and 1.0 ± 0.049 nM for C1–13), making them promising reagents for further test development.Key words: plasmodium, Plasmodium falciparum, malaria, diagnostics, rapid diagnostic test, monoclonal antibodies, glutamate rich protein, dihydrofolate reductase-thymidylate synthase, heme detoxification protein 相似文献
14.
Molecular characterization of a supratypic cross-reactive idiotype associated with IgM autoantibodies 总被引:5,自引:0,他引:5
L F Pratt R Szubin D A Carson T J Kipps 《Journal of immunology (Baltimore, Md. : 1950)》1991,147(6):2041-2046
Neoplastic B cells from patients with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphomas (SLL) frequently express surface Ig reactive with the mouse mAb, Lc1. Raised against a human monoclonal IgM with rheumatoid factor activity, Lc1 detects a major cross-reactive Id (CRI) present on the H chain of many monoclonal IgM autoantibodies. In contrast to other major autoantibody-CRI investigated to date, we note that the Lc1-CRI is expressed by subpopulation of cells in the germinal centers, as well as in the mantle zones, of secondary human B cell follicles. To examine the molecular basis for Lc1 expression, we used the polymerase chain reaction to isolate the functionally rearranged Ig VH genes of monoclonal Lc1-reactive B cell populations from six unrelated patients with CLL or SLL. Although the neoplastic B cells from most patients with CLL or SLL express the CD5 surface differentiation Ag, the lymphoma cells from one patient with SLL were CD5-negative. We find that the Lc1-reactive cells from each cell population have Ig rearrangements involving a VH gene of the VH4 subgroup. However, the VH4 genes rearranged in different Lc1-reactive tumor populations may originate from at least two disparate germ-line VH4 genes. Also, in contrast to the CD5-positive tumor populations, we find evidence for intraclonal diversity in the functionally rearranged VH4 genes of the CD5-negative SLL. Collectively, this study discerns a degeneracy in the VH4 genes that can encode the Lc1 CRI, indicating the term "supratypic cross-reactive idiotype" may best describe the specificity of the Lc1 mAb. Also, this study suggests that expression of CD5 may delineate categories of B cell SLL that differ in their relative rates of constitutive Ig V gene somatic mutation. 相似文献
15.
Mahiuddin Ahmed Ming Cheng Qi Zhao Yehuda Goldgur Sarah M. Cheal Hong-Fen Guo Steven M. Larson Nai-Kong V. Cheung 《The Journal of biological chemistry》2015,290(50):30018-30029
B7-H3 (CD276) is both an inhibitory ligand for natural killer cells and T cells and a tumor antigen that is widely expressed among human solid tumors. Anti-B7-H3 mouse monoclonal antibody 8H9 has been successfully used for radioimmunotherapy for patients with B7-H3(+) tumors. We present the humanization, affinity maturation, and epitope mapping of 8H9 based on structure determination, modeling, and yeast display methods. The crystal structure of ch8H9 Fab fragment was solved to 2.5-Å resolution and used as a template for humanization. By displaying the humanized 8H9 single chain Fv (scFv) on the surface of yeast, the affinity was matured by sequential random mutagenesis and fluorescence-activated cell sorting. Six mutations (three in the complementarity-determining region and three in the framework regions) were identified and incorporated into an affinity-matured humanized 8H9 construct (hu8H9-6m) and an affinity-matured chimeric 8H9 construct (ch8H9-6m). The hu8H9-6m scFv had a 160-fold improvement in affinity (0.9 nm
KD) compared with parental hu8H9 scFv (144 nm
KD). The IgG formats of ch8H9-6m and hu8H9-6m (nanomolar to subnanomolar KD) had 2–9-fold enhancements in affinity compared with their parental forms, potent in vitro antibody-dependent cell-mediated cytotoxicity (0.1–0.3 μg/ml EC50), and high tumor uptake in mouse xenografts. Based on in silico docking studies and experimental validation, the molecular epitope of 8H9 was determined to be dependent on the FG loop of B7-H3, a region critical to its function in immunologic blockade and unique among anti-B7-H3 antibodies published to date. 相似文献
16.
Régis Sodoyer Brigitte Kahn-Perles Torn Strachan Joséphine Sire Marie-José Santoni Corine Layet Pierre Ferrier Bertrand R. Jordan Francois A. Lemonnier 《Immunogenetics》1986,23(4):246-251
The individual contributions of the first two external domains of the HLA-B7 heavy chain to the expression of allele-specific (B7) and locus-specific (B and C) antigenic determinants were investigated using hybrid class I genes. Hybrid genes were constructed in vitro by exon shuffling between the parent genes HLA-B7, HLA-Cw3, HLA-A3, and H-2K
d, and their expression was monitored following transfection into mouse L cells. The results show that most allele-specific antigenic determinants are associated with the first external domain of the 137 heavy chain, whereas all the locus-specific antigenic determinants tested map to the second external domain.Abbreviations used in this paper BSA
bovine serum albumin
- FCS
fetal calf serum
- mAb
monoclonal antibody
- PBL
peripheral blood lymphocytes
- PBS
phosphate-buffered saline 相似文献
17.
The concentration-dependence of the diffusion and sedimentation coefficients (kD and ks, respectively) of a protein can be used to determine the second virial coefficient (B2), a parameter valuable in predicting protein-protein interactions. Accurate measurement of B2 under physiologically and pharmaceutically relevant conditions, however, requires independent measurement of kD and ks via orthogonal techniques. We demonstrate this by utilizing sedimentation velocity (SV) and dynamic light scattering (DLS) to analyze solutions of hen-egg white lysozyme (HEWL) and a monoclonal antibody (mAb1) in different salt solutions. The accuracy of the SV-DLS method was established by comparing measured and literature B2 values for HEWL. In contrast to the assumptions necessary for determining kD and ks via SV alone, kD and ks were of comparable magnitudes, and solution conditions were noted for both HEWL and mAb1 under which 1), kD and ks assumed opposite signs; and 2), kD ≥ ks. Further, we demonstrate the utility of kD and ks as qualitative predictors of protein aggregation through agitation and accelerated stability studies. Aggregation of mAb1 correlated well with B2, kD, and ks, thus establishing the potential for kD to serve as a high-throughput predictor of protein aggregation. 相似文献
18.
Identification of monoclonal antibody defined epitopes on human leukocyte antigens utilizing phage display peptide libraries 总被引:1,自引:0,他引:1
Qinwei Zhou Smruti A. Desai Xinhui Wang Elvyra J. Noronha Mariangela Neri Soldano Ferrone 《Letters in Peptide Science》1999,6(1):77-86
Utilizing phage display peptide libraries, we have identified and mapped the antigenic determinants recognized by mouse monoclonal antibodies (mAb) on two sets of immunologically important molecules, HLA class I and class II antigens. Anti-HLA class I mAb TP25.99 recognizes a conformational and a linear determinant on distinct regions of the HLA class I 3 domain. Anti-HLA class I mAb HO-4 recognizes a conformational determinant on the 2 domain of HLA-A2 and A28 allospecificities. Anti-HLA-DR1, -DR4, -DR6, -DR8, -DR9 mAb SM/549 recognizes a conformational determinant on the chain of HLA class II antigens. These results indicate the versatility of phage display peptide libraries to characterize antigenic determinants recognized by anti-HLA mAb. 相似文献
19.
Hale AH 《Immunogenetics》1980,10(5):469-479
With the use of monospecific rabbit anti-G protein and mouse monoclonal anti-H-2Kk, we have analyzed the spatial relationship of the serologically defined H-2Kk antigens and the major surface glycoprotein (G protein) of vesicular stomatitis virus (VSV) to those antigens recognized by B10.A (k, d) anti-VSV cytotoxic T lymphocytes (CTLs). The ability of monoclonal anti-H-2Kk or rabbit anti-G protein to inhibit specifically the cytolytic activity of B10.A anti-VSV CTLs indicates that the G protein and the H-2Kk molecules are in close proximity to the viral and H-2Kk antigens recognized by the anti-VSV (CTLs. By the method of sequential immunoprecipitation, we also demonstrated that only 10–30 percent of the serologically defined G and H-2Kk molecules are in theG-H-2K
k
complexes.Abbreviations used in this paper Con A
Concanavalin A
- cpm
counts per minure
- CTLs
cytotoxic T lymphocytes
- E: T ratio
effector: target ratio
- G
major surface glycoprotein of VSV
- MHC
major histocompatibility complex
- MOI
multiplicity of infection
- NP40
Nonidet-P40
- PAGE
polyacrylamide gel electrophoresis
- PBS
phosphate-buffered saline
- SaCI
Staphylococcus aureus, Cowan I strain
- SDS
sodium dodecyl sulfate
- UV
ultraviolet light 相似文献
20.
Limeng Zhang Hua Zhang Ziyao Fan Xue Zhou Liquan Yu Hunan Sun Zhijun Wu Yongzhong Yu Baifen Song Jinzhu Ma Chunyu Tong Xintong Wang Zhanbo Zhu Yudong Cui 《PloS one》2015,10(6)
The GapC of Streptococcus dysgalactiae (S. dysgalactiae) is a highly conserved surface protein that can induce protective humoral immune response in animals. However, B-cell epitopes on the S. dysgalactiae GapC have not been well identified. In this study, a monoclonal antibody (mAb5B7) against the GapC1-150 protein was prepared. After passive transfer, mAb5B7 could partially protect mice against S. dysgalactiae infection. Eleven positive phage clones recognized by mAb5B7 were identified by screening phage-displayed random 12-peptide library, most of which matched the consensus motif DTTQGRFD. The motif sequence exactly matches amino acids 48-55 of the S. dysgalactiae GapC protein. In addition, the motif 48DTTQGRFD55 shows high homology among various streptococcus species. Site-directed mutagenic analysis further confirmed that residues D48, T50, Q51, G52 and F54 formed the core motif of 48DTTQGRFD55. This motif was the minimal determinant of the B-cell epitope recognized by the mAb5B7. As expected, epitope-peptide evoked protective immune response against S. dysgalactiae infection in immunized mice. Taken together, this identified conserved B-cell epitope within S. dysgalactiae GapC could provide very valuable insights for vaccine design against S. dysgalactiae infection. 相似文献