Clonal Analysis of a Human Antibody Response. III. Nucleotide Sequences of Monoclonal IgM, IgG, and IgA to Rabies Virus Reveal Restricted Vκ Gene Utilization, Junctional VκJκ and VλJλ Diversity, and Somatic Hypermutation

AbstractIn previous work, we generated four IgM, five IgG1, and one IgA1 mAbs to rabies virus using B cells from four subjects vaccinated with inactivated rabies virus, a thymus-dependent (TD) mosaic Ag, and sequenced the mAb VHDJH genes. Here, we have cloned the VκJκ and VλJλ genes to complete the primary structure of the Ag-binding site of these mAbs. While the anti-rabies virus mAb selection of Vλ genes (2e.2.2 twice, DPL11, and DPL23) reflected the representation of the Vλ genes in the human haploid genome (stochastic utilization), that of Vκ genes (O2/O12 twice, O8/O18, A3/A19, A27, and L2) did not (p = 0.0018) (nonstochastic utilization). Furthermore, the selection of both Vκ and Vλ genes by the anti-rabies virus mAbs vastly overlapped with that of 557 assorted VκJκ rearrangements, that of 253 VλJλ rearrangements in λ-type gammopathies, and that of other Abs to thymus-dependent Ags, including 23 anti-HIV mAbs and 51 rheumatoid factors, but differed from that of 43 Abs to Haemophilus influenzae type b polysaccharide, a prototypic thymus-independent (TI) Ag. The anti-rabies virus mAb VκJκ and VλJλ segments displayed variable numbers of somatic mutations, which, in mAb58 and the virus-neutralizing mAb57, entailed a significant concentration of amino acid replacements in the complementarity-determining regions (p = 0.0028 and p = 0.0023, respectively), suggesting a selection by Ag. This Ag-dependent somatic selection process was superimposed on a somatic diversification process that occurred at the stage of B cell receptor for Ag rearrangement, and that entailed V gene 3′ truncation and N nucleotide additions to yield heterogeneous CDR3s.</jats:p

Vascular endothelial growth factor expression in non-small-cell lung cancer: Prognostic significance in squamous cell carcinoma

AbstractBackground: Recently, some studies have focused on the tumor angiogenesis and its prognostic value. We studied the expression of vascular endothelial growth factor, microvessel counts, and serum concentrations of vascular endothelial growth factor to investigate their association with clinicopathologic factors and prognosis in non-small-cell lung cancer. Methods: The expression of vascular endothelial growth factor was determined by an immunohistochemical analysis from 91 paraffin specimens of completely resected non-small-cell lung cancers using anti–growth factor polyclonal antibody. Microvessel staining was performed by immunohistochemical analysis with anti–factor VIII–related antigen polyclonal antibody. Measurement of the serum concentrations of vascular endothelial growth factor used the sandwich enzyme-linked immunosorbent assay technique. Results: Expression of vascular endothelial growth factor was detected in 48 of the 91 tumors. The positive ratio was significantly higher in patients with adenocarcinoma than in those with squamous cell carcinoma. The microvessel counts were significantly higher in the patients with nodal metastasis than in those without nodal metastasis. The serum concentrations of vascular endothelial growth factor were also significantly higher in the patients with T3-4 disease than in those with T1-2 disease. The microvessel counts were closely associated with expression of vascular endothelial growth factor. The prognosis of patients with a positive growth factor ratio was significantly worse than that of the patients with a negative ratio (p = 0.002), especially in squamous cell carcinoma. According to a multivariate analysis, only nodal status and expression of vascular endothelial growth factor were found to be independent prognostic factors. Conclusions: The expression of vascular endothelial growth factor was one of the most important prognostic factors in completely resected non-small-cell lung cancer, especially in squamous cell carcinoma. (J Thorac Cardiovasc Surg 1998;115:1007-14