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DTIC ADA621368: Antibodies Expressed by Intratumoral B Cells as the... |
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by Defense Technical Information Center |
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As described in our Annual Report, our initial work |
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focused on the expression, purification, and |
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characterization of monoclonal antibodies (mAbs) cloned |
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from intratumoral B lymphocytes (ITLs). While this work |
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was successful in identifying a mAb with preferential |
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binding to tropomyosin 4, all cloned mAbs exhibited |
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polyreactivity. This suggested that the isolated ITLs |
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that we used for mAb production were not members of |
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clonal B cell populations arising in response to |
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stimulation by intratumoral antigens. Hence, our most |
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recent efforts have focused on methods to permit the |
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cloning and expression of recombinant antibodies |
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specifically from oligoclonal ITLs. We accomplished this |
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by performing deep sequencing of VH and VL Ig genes from |
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ITLs from seven additional lung cancer patients, We |
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identified a total of 155,901 functional VH gene |
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sequences with individual patient samples containing from |
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3,327 to 77,182 functional sequences. Although we |
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obtained evidence for somatic hypermutation among these |
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sequences, we did not identify any dominant clones. |
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Sequences representing all VH gene families were |
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identified and exhibited a distribution typically found |
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in peripheral B cells. This suggests that it may be |
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necessary to sequence Ig genes from microdissected |
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intratumoral germinal centers in order to identify clonal |
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ITL populations. |
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Date Published: 2018-11-02 01:32:15 |
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Identifier: DTIC_ADA621368 |
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Item Size: 7932767 |
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Language: english |
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Media Type: texts |
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DTIC Archive; ; DUKE UNIV DURHAM NC; ... |
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