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To improve the sensitivity of a kit, ESPLINE INFLUENZA A&B for rapid diagnosis of influenza by detecting influenza A or B virus specific nucleoproteins (NP), the ESPLINE INFLUENZA A&B-N was developed by using newly established monoclonal antibodies (MAbs) to the respective NP molecule. MAbs FVA2-11 and FrB1-03 recognize the epitope on the amino acid region 59-130aa of the NP molecule of influenza A virus, and that on the region 72-191aa of the NP of influenza B virus, respectively. The new kit detected influenza A and B virus antigens with a detection limit of 10(2.0)-10 (2.7) pfu/test, which is 4-1000 times higher than that of the original kit. Importantly, this kit detected each of influenza A viruses of the known hemagglutinin (HA) subtypes (H1-H15) including the H5N1 viruses recently isolated from human and avian sources in Asia. The kit also detected all of the 15 representative influenza B virus strains tested. The ESPLINE INFLUENZA A&B-N is thus a rapid and highly sensitive and specific kit for the diagnosis of either influenza A or B virus infections.

Immunization with Rous sarcoma virus (RSV)-induced mouse tumors or with SR-3Y1 or NY8-3Y1 rat fibroblasts transformed by a wild type RSV or an env gene deletion mutant of RSV induced complete transplantation resistance against an RSV-induced mouse tumor (CSA1M) in syngeneic hosts. On the other hand, 10 of 19 mice immunized with ts68-3Y1 rat fibroblasts transformed by an src gene temperature-sensitive mutant of RSV (permissive temperature: 33–35° C ) could not reject the CSA1M. SR-3Y1, NY8-3Y1 and RSV-induced mouse tumors expressed a common tumor-specific cell-surface antigen (TSSA) detected by a syngeneic rat antiserum against NY8-3Y1. In ts68-3Y1, expression of the TSSA was temperature-sensitive, TSSA being detected only when ts68-3Y1 was cultivated at the permissive temperature. Immunoprecipitation showed that serum from a rabbit bearing an RSV-induced tumor detected a 60- kdalton protein in cell extracts from SR-3Y1 and NY8-3Y1. Antiserum to NY8-3Y1 failed to detect this protein. These results suggest that the tumor-specific surface antigen(s) on RSV-induced mammalian tumors was coded for by an RSV src gene that was not identical with the simple form of the 60- kdalton protein identified.

Hybrid cell lines were prepared by the fusion of BALB/c myeloma P3U-1 cells with the lymphocytes of BALB/c mice that were immunized with syngeneic Rous sarcoma virus (RSV)-induced tumor CSA1M cells. Three clones of the hybrid progeny (3.4B2, 3.4C6, and 3.5C11) produced cytotoxic IgM antibodies against CSA1M cells. One of the clones, 3.5C11, was chosen for analysis of the detailed specificity. Both direct cytotoxicity assays and absorption tests revealed that monoclonal antibody from 3.5C11 was positive only with CSA1M cells and that it failed to react with other tumors, including 20 RSV-induced mouse tumors, and normal cells. The 3.5C11 monoclonal antibody alone, with or without exogenous complement, was suppressive in the therapy of ip injected CSA1M tumor in syngeneic hosts, and significant prolongation in survival was seen in the treated mice. These results clearly showed presence of an individually distinct tumor-specific cell surface antigen on an RSV-induced mouse tumor.