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Biomedical subjects

F Milgrom

Publications and source records attributed to F Milgrom.

At least 19 recordsLinked to original sources

Studies on avian spinal cord chimeras. I. Neurological and histological manifestations.

Spinal cord chimeras were produced by replacing a small fragment of neural tube of a 2-day-old White Leghorn chicken embryo with a similar fragment from a Japanese quail embryo. The embryo mortality was 61%, and 72% of hatched birds were 'cripples' and had to be sacrificed within 5 days after hatching. Forty-nine chimeras, 10.9% of the total number of operated embryos, were alive for more than 3 weeks. For at least 17 days after hatching, all birds behaved like normal chicks, and the grey quail-like feathers were the only manifestations of their chimerism. Initial neurological symptoms of unsteady walking and drooping of the wings were noted in all birds except for 1 that died an accidental death before it became sick. Advanced symptoms characterized by paralysis of the legs forcing the bird to lie on its side were noted in 40 birds. The chimeras could be divided into two groups, each consisting of 24 birds. The short-survival (SS) chimeras of the first group became terminally ill and had to be sacrificed within 3 months. The long-survival (LS) chimeras of the second group showed more protracted disease, in that only 16 of them showed symptoms of the advanced disease, and the majority showed partial or complete recovery. Ten of the LS birds were kept alive for more than 8 months. Furthermore, many LS chimeras lost their grey feathers. The hallmarks of neurohistological manifestations were mononuclear cell infiltrates, demyelinization with preservation of axons and scar formation. These lesions were restricted to the quail fragment of the spinal cord except for 2 birds in which distant cellular infiltrates were observed. Direct immunofluorescence tests for chicken IgG were positive in spinal cords of most SS chimeras but only of some LS chimeras.

Animals

Studies on avian spinal cord chimeras. II. Immune response of the chicken host to the graft of quail tissue.

A previous study described clinical and pathological manifestations observed in 49 chimeras produced by replacing a small fragment of the neural tube of a chicken embryo by a similar fragment from a Japanese quail embryo. Predominant antibodies in sera of these birds were directed against xenogeneic antigens which are devoid of organ specificity and which are present in quail tissues but absent from chicken tissues. Mixed agglutination test with quail cell monolayers detected such antibodies in sera of all chimeras tested. In most instances, positive reactions were observed already in the 4th week after hatching; they persisted in all birds throughout the observation period of up to 8 months. Some of the detected antibodies were directed against saline-nonextractable surface antigens of quail cells. Enzyme immunoassay with quail organ extracts, agglutination of quail tissue particles, and indirect immunofluorescence test with quail organ sections were positive with sera of many, but not all, chimeras. CNS-specific antibodies, apparently autoantibodies, were detected in serum samples of only one chimera, using enzyme immunoassay with extract of chicken brain and indirect immunofluorescence test with sections of chicken spinal cord. Eluates from lesional spinal cord contained antibodies to non-organ-specific quail antigens but not to CNS-specific antigens. Cerebrospinal fluids of many chimeras had antibodies to quail antigens, but no evidence for antibody formation within the CNS was obtained. Cell-mediated immunity could be demonstrated in all chimeras tested by means of lymphocyte proliferation test after stimulation by quail organ extract. It was concluded that pathological events in the studied chimeras have been most likely mediated primarily by humoral immune responses to non-organ-specific quail antigens.

Animals

Antibodies for trypsinized human red blood cells in human sera.

During studies of agglutination of trypsinized human red blood cells (RBC) by anti-Rh sera, it was noticed that some human sera without Rh antibodies agglutinated these erythrocytes. Of 933 normal and pathological sera subsequently screened, 116 produced such agglutination. The agglutinins were of IgM nature and were directed against an antigen exposed or created by tryptic digestion of RBC. They reacted equally well with autologous as with allogeneic trypsinized RBC and were different from T agglutinins and from Paul-Bunnell antibodies. The mode of formation of these antibodies remains unknown.

Antibodies

Glomerular lesions induced in the rabbit by physicochemically altered homologous IgG.

Immunization of rabbits with physicochemically altered homologous or even autologous IgG induces formation of antibodies combining with IgG of rabbit and of foreign species. Cardiac but not renal lesions were reported in such animals. This study examined the nephritogenic potential of the immune response to cationized or heat-aggregated homologous IgG of b9 or b4 allotype in rabbits of the b4 allotype. Rabbits injected with either b9 or b4 cationized IgG produced antibodies reactive with rabbit and human IgG and with histones; they also developed abnormal glomerular deposits of IgG b4 and C3 corresponding to alterations of the glomerular basement membranes (GBM). Rabbits injected with either b9 or b4 aggregated IgG developed antibodies reactive with rabbit and human IgG and abnormal glomerular deposits of IgG b4 and C3 in the GBM and in the mesangium with subendothelial and mesangial electron-dense deposits. Some rabbits in both groups had proliferative and exudative glomerulonephritis and proteinuria. The results showed that immunization of rabbits with physicochemically altered homologous IgG induces an immune response to rabbit and human IgG and to histones as well as glomerular deposits of autologous IgG and C3 and other glomerular lesions.

Animals

Humoral transplantation antibodies play a role in protracted rejection of murine renal allografts.

Murine renal allografts were studied using (C57BL/6J x A/J)F1 mice as recipients and DBA/2 mice as donors. In this strain combination, protracted rejection was noted in that the circulation was maintained in the graft for over 10 weeks. In all grafts examined after 3 weeks, mononuclear cell infiltrates were noted; in addition, all grafts had immune deposits, apparently containing transplantation antibodies, in glomeruli, tubuli and vessels. These results stressed the role of humoral immunity in protracted renal allograft rejection.

Animals

Glomerulonephritis in NZW kidneys grafted into NZB/W mice.

After left nephrectomy, 3 10-week old NZB/W mice received orthotopic grafts of kidneys from parental NZW mice of the same age. At autopsy conducted at the age of 33-38 weeks, glomerulonephritis of similar extent was noted in the recipients' own and in the grafted kidneys. Also, very similar granular deposits of immunoglobulins and complement were demonstrated in these kidneys. It was concluded that the absence of glomerulonephritis in NZW mice cannot be attributed to the refractoriness of their kidney to this disease.

Animals

Studies on immunological tolerance induced in mice by kidney allografts.

(C57BL/6 x A/J)F1 murine recipients of DBA/2 kidney allografts developed tolerance to DBA/2 tissues, which was measured by observation of growth of a DBA/2 tumor. Eleven, 14 and 18 days after inoculation, the size of the tumor was considerably larger in kidney-grafted than in nongrafted animals. Still, the susceptibility of grafted animals to the tumor did not equal the susceptibility of the DBA/2 mice syngeneic with the tumor. Removal of the renal graft left the recipients with significant tolerance which, however, was weaker than that of the mice in which the kidney graft was left undisturbed. In parabiosis experiments, it was noted that the size of the DBA/2 tumor was equal in the partner which received the DBA/2 kidney graft and in the partner which was not grafted. These experiments rather clearly showed that the tolerance studied was of an 'infectious' type and that it was apparently transferred by some suppressing factor(s) present in the circulation.

Animals

Detection of antigens from gram-negative bacilli in urine of children with urinary tract infections. I. Common antigen of Enterobacteriaceae.

Enterobacterial common antigen (ECA) has attracted considerable interest since the original publication by Kunin in 1962. In the present study we demonstrated this antigen directly in the urine from patients with urinary tract infections (UTI) elicited by enterobacteria. Sheep erythrocytes were incubated with UTI urine; this resulted in their coating with ECA, which was studied by means of hemagglutination by anti-ECA serum. Test tube hemagglutination and the more simple slide hemagglutination were employed and with both procedures similar results were obtained. Positive results were observed in 94-99% of urine specimens from enterobacterial UTI collected in The Buffalo Children's Hospital in the 1960s. ECA in urine could also be demonstrated by hemagglutination inhibition. In this test, antibodies in anti-ECA serum were neutralized as a result of incubation of this serum with urine, and agglutination by the antiserum of sheep erythrocytes coated with a standard ECA preparation was prevented or reduced. By means of this latter test, ECA could be demonstrated in 67-88% of urine specimens from enterobacterial UTI. The possible diagnostic application of these tests has been discussed.

Antigens, Bacterial

Detection of antigens from gram-negative bacilli in urine of children with urinary tract infections. II. Thermostable bacterial proteins.

An enzyme immunoassay for detection of bacterial antigens in urine specimens from urinary tract infections (UTI) was developed. The antigens detected in infections by Enterobacteriaceae: Escherichia coli, Enterobacter aerogenes, and Proteus sp. were different from the enterobacterial common antigen (ECA). They were digested by trypsin and were characterized by remarkable thermostability; their molecular weights were approximately 34,000 and 31,000. They were detected by means of antisera to ECA or to suspension of E. coli. The frequency of positive results with urine specimens from enterobacterial UTI was 87-99%. In contrast, the urine specimens of UTI by Pseudomonas aeruginosa were positive in fewer than 20% with anti-E. coli sera, but were positive in more than 85% in tests with antiserum to P. aeruginosa. The possible diagnostic application of the described tests was discussed.

Antigens, Bacterial

Unusual precipitins in pathological human sera.

In conducting double diffusion gel precipitation tests, unexpected, strong precipitation reactions were noticed between a serum originating from a renal graft recipient and many pathological human sera. The highest frequency of positive reactions were produced by sera of patients with infectious mononucleosis (83%), chronic Epstein-Barr virus infections (72%), rheumatoid arthritis (57%), lepromatous leprosy (57%), and AIDS (44%). The precipitin in these pathological sera was identified as an antibody of IgM variety and the precipitinogen in the transplantation serum was shown to be a thermostable component with beta-globulin mobility. No explanation of the nature of reactions observed can be given at present.

Acquired Immunodeficiency Syndrome

IgG rheumatoid factor in human and rabbit transplantation sera.

Sera of human recipients of kidney allografts and rabbit recipients of skin allografts were tested by means of enzyme immunoassays (EIA) for IgG rheumatoid factor (RF) combining with human IgG (RF-H) or rabbit IgG (RF-R). In humans, the mean OD value in EIA for RF-H was significantly higher in 185 sera of allograft recipients than in 46 pretransplantation sera and 100 sera of normal subjects. Furthermore, the mean OD in patients who rejected their grafts was higher than in patients with satisfactory graft function. Interestingly, there was no difference in OD values for RF-R between sera of graft recipients and sera of normal subjects. Of 52 rabbit recipients of skin allografts, 24 were positive for both RF-R and RF-H, 4 were positive only for RF-R, another 4 only for RF-H, and 20 were negative for both. Renal allografts originating from the previous donors of skin grafts resulted, in most instances, in a significant decrease in the strength of the reaction for both RF-R and RF-H. The possible role of graft rejection in stimulation of RF formation as well as the possible role of RF in graft rejection should be considered.

Animals

Multispecificity of monoclonal rheumatoid factor.

The study involved a monoclonal rheumatoid factor referred to as RF-AN, obtained by Steinitz and his associates from IgG-reactive human lymphocytes 'immortalized' by infection with Epstein-Barr virus. RF-AN combined with red blood cells (RBC) sensitized by either human or rabbit IgG antibodies. The monoclonal character of RF-AN strongly suggested that the same molecule of this factor combined with IgG of both species. Additional evidence for this contention was obtained from absorption and mixed agglutination experiments. RBC sensitized by either human or rabbit antibodies would remove serological activity of RF-AN for both human and rabbit IgG. RF-AN produced exclusively mixed agglutinates when reacted with a mixture of human RBC sensitized by human antibodies and chicken RBC sensitized by rabbit antibodies. Furthermore, it was shown that inhibition with aggregated human Fraction II gave strictly 'specific' results in that it abolished the reaction of RF-AN with RBC sensitized by human antibodies, but not by rabbit antibodies. This result was interpreted as indicating that RF-AN has a multispecific character, i.e., has separate combining sites for human and rabbit IgG. Interestingly, specific inhibition could not be achieved with aggregated rabbit Fraction II and this preparation affected reactivity of RF-AN with RBC sensitized by human antibodies as well as by rabbit antibodies.

Animals

Serological relationship between rat and human carcinoembryonic antigen.

Enzyme immunoassays showed that a rabbit antiserum to rat carcinoembryonic antigen (CEA) had high activity with extracts containing human CEA, which was almost completely inhibited by extracts containing human or rat CEA. Little or no inhibition was obtained with extracts of normal human or rat tissues or by human blood group A or B substances. A monkey antiserum to human CEA had strong activity against extracts containing rat CEA, and little or no activity against extracts of normal rat liver or kidney or a CEA-negative rat colon tumor. Activity of the monkey antiserum against rat CEA was almost completely inhibited by extracts containing rat or human CEA. Little or no inhibition was obtained with extracts of normal rat or human tissues. These results suggest that human and rat CEA share antigenic determinants and that they may be analogous moieties.

Animals

Development of rheumatoid factor research through 50 years.

The discovery of Waaler in 1937 initiated fruitful research on RFs. It was not until the early 1960s that investigators in the field agreed that RFs are antibodies to Fc fragment of IgG. Separate factors combining with human and rabbit IgG and a factor combining with both these IgGs were at first demonstrated by mixed agglutination and then by separation through IgG-conjugated columns. Only RF combining with native autologous IgG should be considered an autoantibody. Other RFs are allo- or heteroantibodies. Cross-reacting RFs that combine with IgG of various species or with IgG and antigens of cell nuclei are of considerable interest. This cross-reactivity may be due to a combining site interacting with shared epitopes or otherwise to multispecificity of the RF molecule in that it has separate or partially overlapping combining sites acting on different epitopes. Experimental studies conducted since the mid-1950s showed that formation of RFs may be elicited by altered autologous IgG. Under natural conditions such alteration was shown to result from interaction of IgG antibody with its corresponding antigen and RF in many infectious diseases and possibly also in rheumatoid arthritis appeared to result from stimulation by immune complexes. More recently alterations of IgG by its reaction with microbial Fc receptors as well as non-specific polyclonal stimulation of B cells were shown to play a role in RF formation. RFs have been implemented in the pathogenicity of rheumatoid arthritis. Recent studies on dispersion of immune complexes in tissue sections by aggregated IgG showed that self-polymerization of IgG RFs results in formation of glomerular deposits in various nephropathies.

Antibodies, Anti-Idiotypic

Induction in human allograft recipients of unresponsiveness to anti-lymphocyte globulin.

The observations presented here confirm previous reports that polyclonal ALG prepared at the University of Minnesota or ATGAM of The Upjohn Co., administered as described, rarely induced sensitization of patients to the horse gamma globulin. In addition, the phenomena of transient antibody production prior to the onset of unresponsiveness and the induction of unresponsiveness in individuals with preexisting antibodies were observed.

Animals

A nonimmunoglobulin precipitin to tissue extracts in pathological human sera.

An alpha-globulin component was noted in pathological human sera, which produced gel precipitation reactions with extracts of human and animal liver. The highest incidence of the precipitin was found in malaria (95%), renal graft rejection (81%), and rheumatoid arthritis (57%). The precipitinogen was thermostable and ethanol soluble; of two precipitation lines formed by this component, one merged into identity reaction with a line produced by commercial lecithin of bovine origin. The possible diagnostic application of the reactions noted was considered.

Alpha-Globulins