Leonard J. Greenberg

Tumor marker kinetics in the monitoring of breast cancer

Controversy exists in using carcinoembryonic antigen (CEA) for monitoring the clinical course of breast cancer. In this study, the kinetics of two plasma tumor markers, CEA and CA15‐3, immediately after the initiation of chemotherapy were assessed in 30 patients with advanced breast cancer. Four distinct kinetic patterns were seen. Two patterns fitted the expected relationship where the plasma marker increased during tumor progression (nine patients), and declined in tumor regression (five patients). The third pattern was paradoxical in that objective tumor regression in eight patients was associated with an acute surge of these markers followed by a steady decline. The doubling times for both CEA and CA15‐3 were immediately shortened four‐fold after therapy suggesting tumor cytolysis in treatment responders. Equally paradoxical was the fourth pattern where tumor progression in eight patients was associated with a rapid and transient decline of markers followed by rebounds. Such a rapid decline may be due to a suppression of marker release, as demonstrated in an in vitro study. Adequate knowledge of these putative paradoxical patterns will permit their effective use in monitoring the disease course and perhaps in the early prediction of the therapeutic response.

Genetic analysis of patients with chronic active hepatitis.

21 patients with chronic active hapatitis (CAH) and their families were HL-A typed. HL-A8 was significantly increased in frequency. An apparent increased frequency of HL-A1 was shown to be secondary to the increased HL-A8 due to linkage disequilibrium. Genotype analysis revealed a striking increased frequency of homozygosity for HL-A8, 6 of 21 patients (28.5%) vs. 2.8% of controls. Two patients and one normal who were homozygous for both HL-A1 and HL-A8 were found to be homozygous for a mixed lymphocyte culture (MLC) determinant 8a. Homozygous 8a cells were used as test-stimulating cells in one-way MLC reactions to determine the frequency of the expression of the 8a determinant in 17 patients and 49 controls selected for HL-A type. 8a was found to be associated with 50% of HL-A8 haplotypes and was frequent in the patient and control populations of the same HL-A types. These data suggest that susceptibility to CAH is determined by homozygosity for a gene that is in linkage disequilibrium with HL-A8 and more closely associated with the HL-A second locus then with the locus for the major MLC determinant.

Hypogammaglobulinemia in uremic infants receiving peritoneal dialysis.

7. Borkowsky W, Krasinski K, Paul D~ et al. Human immunodeficiency virus infections in infants negative for anti-H1V by enzyme-linked immunoassay. Lancet 1987;1:1168-71. 8. Consortium for Retrovirus Serology Standardization. Serologic diagnosis of human immunodeficiency virus infection by Western blot testing. JAMA 1988;260:674-8. 9. Centers for Disease Control. Classification system for human immunodeficiency virus (HIV) infection in children under 13 years of age. MMWR 1987;36:91-6. l 0. Johnson J, Davis E, Sbinaberry R, Nair P. Serologic responses to HIV infection in infants. Presented at the Fifth International Conference on AIDS, Montreal, Quebec, Canada, June 4-9, 1989. 11. Martin K, Katz B, Miller G. AIDS and antibodies to human immunodeficiency virus in children and their families. J Infect Dis 1987;155:54-9. 12. Epstein L, Boucher C, Morrison S, et al. Persistent human immunodeficiency virus type 1 antigenemia in children correlates with disease progression. Pediatrics 1988;82:919-24. 13. Ou C-Y, Kwok S, Mitchell SW, et al. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science 1988;7:825-35. 14. Amadori A, De Rossi A, Gaguinto C, et al. In vitro production of HIV-specific antibody in children at risk of AIDS. Lancet 1988;1:852-4.

Linkage Analysis between the Major Histocompatibility System and Insulin-dependent Diabetes in Families with Patients in Two Consecutive Generations

We have histocompatibility (HLA) genotyped 28 families with insulin-dependent diabetics in two or more consecutive generations, usually parent and child. This strategy of ascertainment was used to maximize the likelihood of obtaining a homogeneous type of disease within a family, and an autosomal dominant mode of inheritance. 76 diabetics and 169 nondiabetics were studied in these families. The frequencies of the antigens Dw3 and Dw4, and the genotype Dw3/Dw4 among the diabetics are 59, 68, and 30%, respectively, as compared with 15, 12, and 2% in normal controls, and 43, 41, and 10% in the nondiabetic relatives of the diabetics. Dw2 is present in only one diabetic (4%), as compared with 18% in normal controls and 17% in nondiabetic relatives.HLA haplotype concordance was analyzed for sib pairs in relation to the haplotype shared by the affected parent/child pair, and for the diabetic sib pairs within each sibship. The results failed to reveal deviations in the expected HLA haplotype assortment. Assuming an autosomal dominant mode and several penetrance levels, linkage analysis between the HLA and diabetes was performed. The total lod score is 0.37 for a recombination fraction of 0.29 at 50% penetrance. Although the linkage and concordance analysis results are inconclusive, they seem to be different from those reported by us for families with normal parents and two or more diabetic sibs. Because ascertainment biases may have influenced these results in an unquantifiable manner, it is not certain whether the two types of families are genetically different. However, the marked difference in the lod scores for the 50% penetrant autosomal recessive model between the two types of families is compatible with a genetic dissimilarity between them. The high frequency of the Dw3 and Dw4 antigens, the Dw3/Dw4 genotype, and the decreased frequency of Dw2, however, indicate the existence of two or more important diabetic genetic factors associated with the D region of the HLA in these families.

Tumor immunology, autoimmunity and aging.

ABSTRACT: Cell‐mediated immune function declines with aging, and may be associated with autoimmunity and malignancy. Humoral immune responses also decline with aging. The chief age‐related effect on the immune system is a decrease in T‐cell function. The “thymus clock” and immunogenesis are discussed in relation to aging. In animals, attempts at immunologic rejuvenation by cellular or hormonal means have not been as successful as the results attained by genetic manipulation.

Measurement of Precipitin Reactions in the Millimicrogram Protein-Nitrogen Range

Exploitation of newer instrumentation and a dye-binding method for protein measurement made possible reduction in volume down to 1/1000 that commonly used, with no greater error. The procedure was tested at two levels, 1- and 10-�l volumes, with human gamma globulin and rabbit antiserum. Of the dyes tested, bromsulfalein proved best for the protein estimation.

Immunogenetics of response to a purified antigen from group A streptococci. I. Complex segregation analysis.

A purified, extracellular protein, M. W. 17500, from group A streptococci was used in an in vitro model to study genetic aspects of immune responsiveness. Peripheral blood lymphocytes from members of eight nuclear families and one kindred of three generations were isolated and purified by Ficoll-Isopaque flotation and cultured together with varying concentrations of antigen. A summary measure of the antigen response data (measured as CPM) was constructed through a weighted regression analysis of response on dose. Commingling analysis of the standardized regression coefficients provided evidence for the existence of two underlaying distributions. Segregation analysis confirmed that response to this antigen is controlled by a major gene and is inherited in an autosomal dominant manner.

Serological Inhibition of Blast Transformation to Purified Streptococcal Antigens by Planned Immunization in HLA (A, B) Compatible Unrelated Individuals

Abstract. Sera obtained from planned immunizations between unrelated donors and recipients, identical or compatible at HLA‐A and B, were assessed for their capacity to alter the in vitro response of a test panel of lymphocytes to PHA and a purified streptococcal antigen (PSA). In the case of PHA, no serum effects were apparent. The response to PSA, however, significantly inhibited by two sera. When tested for their complement‐dependent cytotoxicity on enriched populations of T and B lymphocytes, none of the sera manifested cytotoxicity against T cells nor did serological inhibition correlate with the capacity to lyze B cells. The data suggest that inhibition of the PSA response is mediated by blocking antibodies specific for a subset of lymphocytes, possibly T cells. While the precise mechanism governing the response to PSA is not known, the data are compatible with the idea that an HLA‐linked Ir gene, expressed on a subset of T lymphocytes, controls immune responsiveness to PSA.

HLA-Linked Regulation of Immune Responsiveness in Man

During the past decade, there has been an ever-increasing interest in the interaction between the immune system and the major histocompatibility complex (MHC) of the species. The impetus for this intense scientific focus derives for the most part from the discovery that immune responsiveness is genetically controlled by genes mapping in the H-2 complex of the mouse, i.e., the observation of linkage between the Ir-1 locus and H-2 (McDevitt and Chinitz, 1969). Since that time, many associations between immune-response (Ir) genes and the MHC have been detected in inbred strains of mice (Martin et al., 1971; Vaz and Levine, 1970; McDevitt et al., 1969) and guinea pigs (Green and Benacerraf, 1971; Ellman et al., 1970; Green et al., 1970). This subject has been extensively reviewed in several books (McDevitt, 1978; Sercarz et al., 1977; Snell et al., 1976; Klein, 1975) and will not be dealt with in any detail in this chapter.

Genetic, Developmental, and Evolutionary Aspects of Life Span

In a broad sense, life span and aging can be defined as the length of survival and the collective changes that occur during the period between conception and death (Buerger, 1957). Like most biological phenomena, there is ample evidence to support both genetic and environmental approaches to the study of life span and aging (Comfort, 1974). Although life, aging, and death are generally accepted as essential to the evolutionary process, little is known about the factors controlling the life span of a species at the phenotypic level. As a consequence, numerous theories have been proposed to explain the aging process. These theories have implicated biological clocks (Landahl, 1959; Burnet, 1973), the waning of immunologic vigor (Ram, 1967; Walford, 1969; Greenberg and Yunis, 1972), the finite lifetime of cells (Strehler, 1966; Novelli, 1970; Hayflick, 1965, 1974), crosslinking of macromolecules (Bjorksten, 1968), somatic mutation (Mole, 1963; Jones and Kimmeldorf, 1964), accumulation of random errors (Orgel, 1963, 1973), dietary caloric intake (Ross, 1959; Mclntyre et al., 1964), and many others.