Antiidiotypic response against murine monoclonal antibodies reactive with tumor-associated antigen TAG-72.

The immune response of 42 gastrointestinal and ovarian cancer patients at 1 month after exposure to murine monoclonal antibodies (B72.3 and CC49) reactive with the tumor-associated antigen TAG-72 was studied. The incidence of human anti-mouse antibody response was 89% to B72.3 and 70% to CC49. To evaluate the antiidiotypic immune response, we developed a serological assay based on affinity chromatography to remove the interference due to the presence of TAG-72, antiisotypic, and antiallotypic immunoglobulins in the serum. Seventy-eight percent of patients who received B72.3 developed an antiidiotypic response; in 33% of the patients, this was the only immune response detected. The antiidiotypic immune response after treatment with CC49 was present in 54% of the patients. Twelve percent of the patients who received CC49 developed an antiidiotypic response in the absence of antiisotypic or antiallotypic immune response. The lower immunogenicity of the variable region of CC49 is encouraging when considering the use of chimeric or humanized antibodies derived from the murine monoclonal antibody CC49 in clinical studies.

Conformationally biased analogs of human C5a mediate changes in vascular permeability.

A panel of conformationally constrained, decapeptide agonists corresponding to the C-terminal "effector" region of human C5a (C5a65-74 or ISHKDMQLGR) was evaluated for the ability to increase vascular permeability. One constrained analog, acyl-YSFKPMPLaR, expressed between 2 and 10% of full C5a activity in increasing vascular permeability, as measured by the extravasation of Evans blue dye in guinea pig skin. This analog was at least 10-fold more potent than its unconstrained sister analog C5a65-74465, F67++ (YSFKDMQLGR), which was used as an internal standard in these assays. Neither acyl-YSFKPMPLaR nor YSFKDMQLGR changed the transvascular equillibrium of an electrolyte, 86Rb, at the peptide injection site. However, both peptides effected a significant increase in the extravasation of two macromolecules, 125I-labeled bovine serum albumin and 131I-labeled monoclonal antibody BL-3. The extravasation of Evans blue dye mediated by 0.03 to 0.1 nmol of acyl-YSFKPMPLaR was nearly abolished by 1 to 10 nmol of the antihistamine diphenhydramine. For YSFKDMQLGR, however, the sensitivity toward diphenhydramine was observed only at low concentrations of the peptide (1 nmol). When incubated in human and mouse sera, acyl-YSFKPMPLaR was shown to be stable toward the actions of serum carboxypeptidases. However, the unconstrained analog YSFKDMQLGR was rapidly converted to the des-Arg form under the same conditions. Taken together, these results support a growing body of evidence that unique topochemical features expressed in conformationally constrained agonist analogs of C5a contribute favorably to their ability to modulate vascular permeability and to their stability in serum.

Oestrogen retards the development of spontaneous thymomas in BUF/Mna rats.

BUF/Mna rats develop spontaneous thymomas with nearly 100% incidence in both sexes. While the thymomas in males develop from around 9 months of age, those in females start from 13-15 months of age. To clarify the mechanism of the delay of thymomagenesis in females, the effect of sex hormones on the development of thymomas was examined after either gonadectomy or oestrogen treatment. Prepubertal ovariectomy accelerated the thymoma development in females, whereas orchiectomy did not affect it. An intraperitoneal injection of oestriol (20 mg) into males at 2 months of age remarkably diminished the thymic weight to about one-tenth of age-matched controls at 16 months of age. These results suggest that oestrogen can actually retard the onset of thymoma in spite of genetic control of its incidence. However, oestrogen did not cause thymic involution when it was injected into rats over 9 months of age. Immunohistochemically, there seemed to be no distinct difference in distribution of oestrogen-receptor-bearing epithelial cells between thymomas and 2- to 3-month-old thymuses. The oestrogen sensitivity of the thymus might be destined to be lost, as the thymic epithelial cells start neoplastic changes with the impairment of oestrogen-receptor function.

Characterization of intrathymic and extrathymic T cell development in spontaneous thymoma Buffalo/Mna rats.

The developmental status of T cell lineages at prethymic, intrathymic and postthymic stages in spontaneous thymoma Buffalo/Mna (BUF/Mna) rats was characterized on the basis of surface phenotypes and some immune responses. The proportion of bone marrow cell populations was shown to be normal in thymoma rats by immunofluorescence flow cytometry (FACS). In spite of a cortex-predominant appearance of thymomas, the proportion of thymocyte populations was phenotypically normal as assessed by FACS. Double immunohistochemical stainings revealed that a substantial number of single-possible (CD4+CD8- or CD4-CD8+) thymocytes existed not only in the narrowed medullary areas but also in the enlarged cortical areas of the thymomas. In lymph nodes, the proportion of T cells increased with age, reaching 88% after 20 months of age. This increase was due mainly to an increase in the CD8+ population but not the CD4+ population, resulting in low CD4/CD8 ratios. An abnormal increase of Thy1+ immature T cells was also observed in the lymph nodes. However, these phenotypic changes in the T cell lineages in the thymoma rats were not so influential as to alter their immunological reactivities, such as the primary antibody response to a T-dependent antigen, the graft-versus-host reaction and the mixed lymphocyte reaction to allo-antigens. These results suggest the possible presence of some altered differentiation pathways for intrathymic and postthymic T cell development in BUF/Mna rats.

Growth-promoting effect of oestriol in a lymphoma lacking oestrogen receptors.

Various doses (1 microgram to 10 mg) of oestriol (E3) were intraperitoneally injected into mice immediately after subcutaneous inoculation of an oestrogen receptor-negative lymphoma cell line (KE-5) established from a spontaneously developed AKR thymic lymphoma. The growth of KE-5 cells was markedly promoted by E3 at the early stage of tumour growth. At this stage, 1 microgram E3 enhanced tumour growth significantly and the maximum effect was obtained with 1 mg E3. Normal female mice showed a higher incidence and shorter latency than males. However, once tumours became palpable, the tumour growth rate appeared to be unaffected. Histological observations using Alcian blue and colloidal iron revealed a marked increase of hyaluronic acid in the subcutaneous connective tissue of the tumour-injection site within 3-5 days after intraperitoneal administration of 1 mg E3. Biochemical analyses showed a rapid and marked increase in skin hyaluronic acid content to over 3 times the control levels (0.25 +/- 0.10 mg g-1 skin) within 3 days of E3 administration. Subcutaneous inoculation of KE-5 cells together with hyaluronic acid (0.2 mg) resulted in markedly enhanced tumour growth, particularly at the early stage. These results suggest that an increase in stromal hyaluronic acid content is the most likely mechanism responsible for the promoting effect of E3 on KE-5 cells.

Increased vascular permeability in the thymus of the autoimmune New Zealand mouse.

The thymus glands of non-autoimmune BDF1 and C3H mice and autoimmune NZB/WF1 mice were studied histologically at intervals ranging from one hour to 60 days after systemic administration of carbon. In NZB/WF1 mice over 9 weeks of age, many circulating carbon-laden macrophages were seen to have penetrated the walls of blood vessels, and to have then entered the thymic parenchyma. Carbon was also taken up by many perivascular macrophages stretched out along blood vessels and by many resident tissue macrophages scattered throughout the thymic parenchyma. In contrast, no carbon was seen at any time in the extravascular tissues of the thymus in BDF1 and C3H mice of comparable age. These results indicate a great increase in the permeability of blood vessels in the thymus of NZB/WF1 mice. This increase in carbon permeability occurs both in the cortex and the medulla, particularly at the corticomedullary junction. There is little increase in the permeability to carbon in NZB/WF1 mice at the age of 4 weeks, suggesting that the increase in vascular permeability begins between the ages of 4 and 9 weeks. The possible role of this greatly increased blood vessel permeability in the thymus on the aetiology and pathogenesis of autoimmune disease is discussed.