Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus
In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences...
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| Veröffentlicht in: | Scientific reports 2017-10, Vol.7 (1), p.12713-15, Article 12713 |
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| creator | Lu, Zhengchun Tallmadge, Rebecca L. Callaway, Heather M. Felippe, M. Julia B. Parker, John S. L. |
| description | In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus. |
| doi_str_mv | 10.1038/s41598-017-12725-5 |
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Julia B. ; Parker, John S. L.</creator><creatorcontrib>Lu, Zhengchun ; Tallmadge, Rebecca L. ; Callaway, Heather M. ; Felippe, M. Julia B. ; Parker, John S. L.</creatorcontrib><description>In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-017-12725-5</identifier><identifier>PMID: 28983085</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>38/1 ; 38/44 ; 38/77 ; 631/250/248 ; 631/61/185 ; 82/80 ; 82/83 ; Antigens ; Expression vectors ; Feline panleukopenia ; Humanities and Social Sciences ; Immunization ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulins ; Immunotherapy ; Leukocytes (mononuclear) ; Light chains ; Lymphocytes B ; Monoclonal antibodies ; mRNA ; multidisciplinary ; Peripheral blood mononuclear cells ; Science ; Science (multidisciplinary) ; T cell receptors</subject><ispartof>Scientific reports, 2017-10, Vol.7 (1), p.12713-15, Article 12713</ispartof><rights>The Author(s) 2017</rights><rights>2017. 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Julia B.</creatorcontrib><creatorcontrib>Parker, John S. L.</creatorcontrib><title>Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus.</description><subject>38/1</subject><subject>38/44</subject><subject>38/77</subject><subject>631/250/248</subject><subject>631/61/185</subject><subject>82/80</subject><subject>82/83</subject><subject>Antigens</subject><subject>Expression vectors</subject><subject>Feline panleukopenia</subject><subject>Humanities and Social Sciences</subject><subject>Immunization</subject><subject>Immunoglobulin A</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulins</subject><subject>Immunotherapy</subject><subject>Leukocytes (mononuclear)</subject><subject>Light chains</subject><subject>Lymphocytes B</subject><subject>Monoclonal antibodies</subject><subject>mRNA</subject><subject>multidisciplinary</subject><subject>Peripheral blood mononuclear cells</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>T cell receptors</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kduK1TAUhosozjDOC3ghAW-8qebYJDfCMHiCQcHDdUjT1T0Z06Q27YbtQ_jMpnZm2ArmJgnr-_9krb-qnhL8kmCmXmVOhFY1JrImVFJRiwfVKcVc1JRR-vDofFKd53yDyxJUc6IfVydUacWwEqfVry_wY4HoANlowyH7jFKPegg-AvLDsMS0C6ldyh0Nnz9e5MJ1aL4G1MEeQhoHiPMqsZvI_4QODSkmF1IxLPTs29QdUB7B-d47NKc7-9HGAMv3NEL0Fu39tOQn1aPehgznt_tZ9e3tm6-X7-urT-8-XF5c1Y5LPtfgOG97LBVrOcdWMsUVJa5tGyrBYtp3klDbYmtxB0R3mHVWiAY3SmorGGFn1evNd1zaATpXmphsMOPkBzsdTLLe_F2J_trs0t6IhmqiZTF4cWswpTLAPJvBZwch2AhpyYZorqRgDRMFff4PepOWqQxnpYRUXDdipehGuSnlPEF__xmCzZq42RI3JXHzJ3Gzip4dt3Evucu3AGwDcinFHUxHb__f9jev4roF</recordid><startdate>20171005</startdate><enddate>20171005</enddate><creator>Lu, Zhengchun</creator><creator>Tallmadge, Rebecca L.</creator><creator>Callaway, Heather M.</creator><creator>Felippe, M. 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Julia B.</au><au>Parker, John S. L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2017-10-05</date><risdate>2017</risdate><volume>7</volume><issue>1</issue><spage>12713</spage><epage>15</epage><pages>12713-15</pages><artnum>12713</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>28983085</pmid><doi>10.1038/s41598-017-12725-5</doi><tpages>15</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 38/1 38/44 38/77 631/250/248 631/61/185 82/80 82/83 Antigens Expression vectors Feline panleukopenia Humanities and Social Sciences Immunization Immunoglobulin A Immunoglobulin G Immunoglobulins Immunotherapy Leukocytes (mononuclear) Light chains Lymphocytes B Monoclonal antibodies mRNA multidisciplinary Peripheral blood mononuclear cells Science Science (multidisciplinary) T cell receptors |
| title | Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus |
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