Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin
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- Inagaki Takashi INAGAKI Takashi
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- Yoshimi Tatsunari YOSHIMI Tatsunari
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- Kobayashi Satoshi [他] KOBAYASHI Satoshi
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- KAWAHARA Masahiro
- Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo
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- NAGAMUNE Teruyuki
- Department of Bioengineering, Graduate School of Engineering, University of Tokyo
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Author(s)
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- Inagaki Takashi INAGAKI Takashi
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- Yoshimi Tatsunari YOSHIMI Tatsunari
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- Kobayashi Satoshi [他] KOBAYASHI Satoshi
- Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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- KAWAHARA Masahiro
- Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo
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- NAGAMUNE Teruyuki
- Department of Bioengineering, Graduate School of Engineering, University of Tokyo
Abstract
Although various in vitro immunization methods to generate antigen-specific antibodies have been described, a highly effective method that can generate high-affinity immunoglobulins has not yet been reported. Herein, we analyzed a cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulins. We identified a combination of T cell-dependent stimuli (IL-4, IL-5, anti-CD38 and anti-CD4O antibodies) plus lipopolysaccharides (LPS) that stimulates antigen-exposed splenocytes in vitro, followed by induction of the cells phenotypically equivalent to germinal center B cells. We also observed that LPS induced high expression levels of mRNA for activation-induced cytidine deaminase. We stimulated antigen-exposed splenocytes, followed by the accumulation of mutations in immunoglobulin genes. From the immunized splenocytes, hybridoma clones secreting antigen-specific immunoglobulins were obtained.
Journal
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- Journal of bioscience and bioengineering
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Journal of bioscience and bioengineering 115(3), 339-345, 2013-03-25
The Society for Biotechnology, Japan
References: 34
-
1
- Tocilizumab inhibits signal transduction mediated by both mIL-6R and sIL-6R, but not by the receptors of other members of IL-6 cytokine family
-
MIHARA M.
Int. Immunopharmacol. 5, 1731-1740, 2005
Cited by (1)
-
2
- The EGFR as a target for anticancer therapy-focus on cetuximab
-
BASELGA J.
Eur. J. Cancer 37(Suppl. 4), S16-S22, 2001
Cited by (1)
-
3
- Surface display of antibodies
-
LITTLE M.
Biotechnol. Adv. 12, 539-555, 1994
Cited by (1)
-
4
- Ribosome display efficiently selects and evolves high-affinity antibodies in vitro from immune libraries
-
HANES J.
Proc. Natl. Acad. Sci. USA 95, 14130-14135, 1998
Cited by (1)
-
5
- Antibody affinity maturation using bacterial surface display
-
DAUGHERTY P. S.
Protein Eng. 11, 825-832, 1998
Cited by (1)
-
6
- Isolation of anti-T cell receptor scFv mutants by yeast surface display
-
KIEKE M. C.
Protein Eng. 10, 1303-1310, 1997
Cited by (1)
-
7
- Isolation of anti-CD22 Fv with high affinity by Fv display on human cells
-
HO M.
Proc. Natl. Acad. Sci. USA 103, 9637-9642, 2006
Cited by (1)
-
8
- In vitro immunization as an adjunct to the production of hybridomas producing antibodies against the lymphokine osteoclast activating factor
-
LUBEN R. A.
Mol. Immunol. 17, 635-639, 1980
Cited by (1)
-
9
- In vitro immunization. Effect of growth and differentiation factors on antigen-specific B cell activation and production of monoclonal antibodies to autologous antigens and weak immunogens
-
BORREBAECK C. A.
J. Immunol. 136, 3710-3715, 1986
Cited by (1)
-
10
- In vitro immunization of mouse spleen cells for the production of monoclonal IgGI antibodies using an antigen-specific T helper cell clone (D.10.G4.1)
-
UTHOFF S.
J. Immunol. Methods 166, 165-175, 1993
Cited by (1)
-
11
- Production of human-human hybridomas secreting antibody to sheep erythrocytes after in vitro immunization
-
STRIKE L. E.
J. Immunol. 132, 1798-1803, 1984
Cited by (1)
-
12
- An improved in vitro immunization procedure for the production of monoclonal antibodies against neural and other antigens
-
BOSS B. D.
Brain Res. 291, 193-196, 1984
Cited by (1)
-
13
- B cell stimulatory factor-1 (interleukin 4) prepares resting murine B cells to secrete IgG1 upon subsequent stimulation with bacterial lipopolysaccharide
-
SNAPPER C. M.
J. Immunol. 139, 10-17, 1987
Cited by (1)
-
14
- Isotype switching in anti-immunoglobulin-activated B lymphoblasts : differential requirements for interleukin 4 and other lymphokines to elicit membrane vs. secreted IgG1
-
PURKERSON J. M.
Eur. J. Immunol. 21, 707-714, 1991
Cited by (1)
-
15
- Activation-induced cytidine deaminase turns on somatic hypermutation in hybridomas
-
MARTIN A.
Nature 415, 802-806, 2002
Cited by (1)
-
16
- Chronic lymphocytic leukemia B cells expressing AID display dissociation between class switch recombination and somatic hypermutation
-
OPPEZZO P.
Blood 101, 4029-4032, 2003
Cited by (1)
-
17
- Interleukin-5 regulates genes involved in B-cell terminal maturation
-
HORIKAWA K.
Immunology 118, 497-508, 2006
Cited by (1)
-
18
- <no title>
-
KABAT E. A.
Sequences of proteins of immunological interest, 1991
Cited by (1)
-
19
- BCL-6 expression during B-cell activation
-
ALLMAN D.
Blood 87, 5257-5268, 1996
Cited by (1)
-
20
- Identification of murine germinal center B cell subsets defined by the expression of surface isotypes and differentiation antigens
-
SHINALL S. M.
J. Immunol 164, 5729-5738, 2000
Cited by (1)
-
21
- Changing responsive-ness to chemokines allows medullary plasmablasts to leave lymph nodes
-
WEHRLI N.
Eur. J. Immunol. 31, 609-616, 2001
Cited by (1)
-
22
- The pattern and distribution of immunoglobulin VH gene mutations in chronic lymphocytic leukemia B cells are consistent with the canonical somatic hypermutation process
-
MESSMER B. T.
Blood 103, 3490-3495, 2004
Cited by (1)
-
23
- Rapid generation of specific antibodies by enhanced homologous recombination
-
SEO H.
Nat. Biotechnol. 23, 731-735, 2005
Cited by (1)
-
24
- Generation and iterative affinity maturation of antibodies in vitro using hypermutating B-cell lines
-
CUMBERS S. J.
Nat. Biotechnol. 20, 1129-1134, 2002
Cited by (1)
-
25
- Identification of murine B cell lines that undergo somatic hypermutation focused to A : T and G : C residues
-
BHATTACHARYA P.
Eur. J. Immunol. 38, 227-239, 2008
Cited by (1)
-
26
- Hybridoma generation by in vitro immunization of murine splenocytes with cytosolic proteins of Chinese hamster ovary (CHO) mitotic cells
-
RAVI M.
Hybridoma (Larchmt) 26, 311-315, 2007
Cited by (1)
-
27
- Production of monoclonal antibodies against Neisseria meningitidis using popliteal lymph nodes and in viva/in vitro immunization : prevalence study of new monoclonal antibodies in greater Sao Paulo, Brazil
-
BELO E. F.
Hybridoma (Larchmt) 26, 302-310, 2007
Cited by (1)
-
28
- Biochemical and physicochemical characterization of mouse B cell growth factor : a lymphokine distinct from interleukin 2
-
FARRAR J. J.
J. Immunol. 131, 1838-1842, 1983
Cited by (1)
-
29
- B cell growth and differentiation factors and mechanism of B cell activation
-
KISHIMOTO T.
Immunol. Rev. 78, 97-118, 1984
Cited by (1)
-
30
- Novel In Vitro Screening System for Monoclonal Antibodies Using Hypermutating Chicken B Cell Library
-
Todo Kagefumi , Miyake Kenji , Magari Masaki , Kanayama Naoki , Ohmori Hitoshi
Journal of Bioscience and Bioengineering 102(5), 478-481, 2006
J-STAGE Cited by (1)
-
31
- Mouse germinal center B cells with the xid mutation retain responsiveness to antimouse CD40 antibodies but diminish IL-5 responsiveness
-
BABA Masashi , KIKUCHI Yuji , MORI Shigeo , KIMOTO Hiroshi , INUI Seiji , SAKAGUCHI Nobuo , INOUE Jun-ichiro , YAMAMOTO Tadashi , TAKEMORI Toshitada , HOWARD Maureen , TAKATSU Kiyoshi
Int. Immunol. 9(10), 1463-1473, 1997
References (70) Cited by (2)
-
32
- IL-4 and anti-CD40 prtect against Fas-mediated B cell apoptosis and induce B cell growth and differentiation
-
NAKANISHI Kenji , MATSUI Kiyoshi , KASHIWAMURA Shin-ichiro , NISHIOKA Yasuhiro , NOMURA Jun , NISHIMURA Yoshiko , SAKAGUCHI Nobuo , YONEHARA Shin , HIGASHINO Kazuya , SHINKA Sohei
Int. Immunol. 8(5), 791-798, 1996
References (50) Cited by (3)
-
33
- Anti-Peptide Antibody Production Elicited by in Vitro Immunization of Human Peripheral Blood Mononuclear Cells
-
TAMURA Takashi , TOMIMATSU Kosuke , KATAKURA Yoshinori , YAMASHITA Makiko , MATSUMOTO Shin-ei , AIBA Yoshihiro , JUNG Yeon Suk , ABE Yoshiichi , FUJIKI Tsukasa , TERUYA Kiichiro , SHIRAHATA Sanetaka
Bioscience, Biotechnology, and Biochemistry 71(12), 2871-2875, 2007-12-23
J-STAGE Ichushi Web References (17) Cited by (3)
-
34
- Production of Human Monoclonal Antibodies against FcεRIα by a Method Combining in Vitro Immunization with Phage Display
-
TOMIMATSU Kosuke , MATSUMOTO Shin-ei , YAMASHITA Makiko , TERUYA Kiichiro , KATAKURA Yoshinori , KABAYAMA Shigeru , SHIRAHATA Sanetaka
Bioscience, Biotechnology, and Biochemistry 73(7), 1465-1469, 2009-07-23
J-STAGE Ichushi Web References (26) Cited by (1)