Guinea pig line 10 hepatocarcinoma model: characterization of monoclonal antibody and in vivo effect of unconjugated antibody and antibody conjugated to diphtheria toxin A chain.

Monoclonal antibodies were raised against the guinea pig line 10 (L10) hepatocarcinoma, and an IgG1-producing hybridoma (D3) was selected for further study. D3 is a true monoclonal antibody as demonstrated by two-dimensional gel electrophoresis. Radioimmunoassays on live cells revealed no cross-reactivity with normal tissues or with the line 1 hepatocarcinoma which was used as a control. Membrane immunofluorescence assays demonstrated similar specificity. Immunoperoxidase staining of cryostat sections of tumor and normal tissues of both adult animals and fetuses showed that the D3 monoclonal antibody reacted primarily with the L10 tumor, but some cross-reactivity with smooth muscle, placenta, fetal skeletal muscle, and fetal liver was also demonstrated. Radioimmunoprecipitation of detergent extracts of iodinated L10 cells showed that the antigen is present on the cell surface as a dimer of Mr 290,000 (unit size, Mr 148,000). Therapy studies with unconjugated D3 antibody demonstrated a minor dose-dependent effect on tumor growth. D3 antibody conjugated to the A chain of diphtheria toxin (10(-7) M) was cytotoxic to 100% of L10 cells in vitro. Animals treated with a single 1-mg i.v. injection of this immunoconjugate on Day 7 following the intradermal injection of 10(5) tumor cells demonstrated a highly significant inhibition of tumor growth compared to control animals and those treated with unconjugated antibody.

The Hawthorne effect in the measurement of adolescent smoking.

It is possible that the process of repeatedly measuring the smoking behaviour of adolescents may very well affect that behaviour. This paper reports a test for the extent of such a "Hawthorne" effect in a longitudinal survey of smoking by English adolescents. The self-reported smoking behaviour of 15-16 year olds who attended schools which had participated in the study for five years was compared with that of 15-16 year olds who attended other schools. The prevalence of smoking was lower in those schools which had been surveyed for five years. A number of possible explanations for this finding are discussed. It is concluded that such a "Hawthorne" effect is unlikely to bias analyses relying on comparisons within the data set. However, they can certainly bias the prevalance estimates obtained from such a study. Thus they provide yet another reason why prevalence estimates from cohorts studied over a period of time must be used with considerable caution.

Monoclonal antibodies to human colorectal tumor-associated antigens: improved elicitation and subclass restriction.

Monoclonal antibodies to tumor-associated antigens (TAA) of human colorectal cancer were elicited using immunosorbents of lectins combined with peripheral protein extracts of xenografted colon adenocarcinoma. This method of immunization was compared with whole cells from surgical specimens and to crude membranes from xenografted tumors. The immunosorbent immunogens were superior to the other immunogens in three ways: (1) the number of hybrids reactive with colon tumor cells or extracts, but not with lymphoid cells or extracts, (2) the number of stable hybrids after cloning, and (3) the number of hybridoma clones reactive with tissue sections of colon tumors, but not normal colonic mucosa. In addition, lectin immunosorbents elicited primarily IgG antibodies, especially IgG3, with almost 50% of the clones of interest reacting to seemingly less immunogenic glycoproteins. The improved elicitation of monoclonal antibodies to TAA by the use of lectin immunosorbents and peripheral protein extracts has considerable potential for generating reagents useful in diagnosis and therapy of human tumors.

Characterization of infection and replication of Mason-Pfizer monkey virus in human cell cultures.

Human cells derived from both normal and neoplastic tissues can be infected by Mason-Pfizer monkey virus (MPMV) without accompanying cytopathology. Infection of cell cultures such as human rhabdomyosarcoma (A204) results in a persistenly infected cell line which can be subcultured over 30 sequential culture passages without significant change in phenotype properties according to reverse, transcriptase (RT), MPMV p27 antigen content, virus particle count and infectivity titre. Productive virus infections were established at relatively low virus particle (VP) input multiplicities (p.i.m.; about 0.06 VP/cell) In A204 cell cultures. At higher p.i.m. (about 600 to 6000 VP/cell) newly synthesized virus was detected within 4 days post infection. Although virus production was cumulative following primary infection, after subculture of infected cultures MPVM production was greater during active cell division. Using synchronization techniques, MPMV replication in persistently infected cultures was found to be cell cycle-dependent. The major internal antigen, p27, was synthesized in G2 and newly synthesized virus particles were released predominantly during mitosis and early G1. Colcemid arrest of cells during mitosis inhibited subsequent MPMV release. Consequently, production of extracellular virus depends upon the progression of cells through the mitotic stage. These data, which provided a basic understanding of the virus-host relationship that occurs in primate cells productively infected with MPMV, were used as a guideline for isolating MPMV-like viruses from experimentally and naturally infected Rhesus monkey.

The Clara cell: an electron microscopy examination of the terminal bronchioles of rats and monkeys following inhalation of hexachlorocyclopentadiene.

The effects of inhalation of hexachlorocyclopentadiene (Hex) up to 14 wk on the terminal bronchioles of rats and monkeys was examined by electronic microscopy. Exposed rats elicited a concentration-related increase in the incidence of electron-lucent inclusions in the bronchiolar Clara cells when compared to controls. The inclusions in the high-concentration (0.2 ppm) group were round and more abundant than the rod-shaped inclusions observed in the intermediate (0.05 ppm) and low-concentration (0.01 ppm) exposure group. No ultrastructural changes were observed that could be attributed to the inhalation of Hex vapor in exposed monkeys. The origin and significance of these inclusions is discussed in light of the literature.