Multiple Sera Research Articles

REVERSAL OF SIGNS AND SYMPTOMS OF ACQUIRED IMMUNODEFICIENCY SYNDROME (AIDS) WITH PREDNISONE

We describe in detail the case of a male infant of a prostitute and IV drug abuser. The child presented at 9 mo of age with severely delayed growth (6.6 kg) and development, recurrent bacterial infections, lymphadenopathy, hepatosplenomegaly (8 cm), persistent oral candidiasis, and chronic pulmonary infiltrates. Immunologic studies showed polyclonal hypergammaglobulinemia and inverted T-cell T4/T8 ratios. Skin tests to Candida and phytohemagglutinin were positive. Lung biopsy showed peribronchiolar lymphocytic infiltrates. Axillary lymph nodes contained obliterated margins with increased plasmacytes centrally. Liver biopsy demonstrated mild ballooning of hepatocytes. Bone marrow biopsy revealed hypercellularity with increased plasmacytes. EB virus was repeatedly isolated from lymph node, bone marrow, and blood lymphocytes, although antibodies to EBV did not appear for another 4 mo. Sera from both mother and child did contain antibody to human T-cell leukemia virus. Multiple sera were negative for antibody to cytomegalovirus and herpes virus. These viruses failed repeatedly to grow from tissue specimens. Initially, the child's course was observed off all treatment. After 8 wks of slow deterioration, he became lethargic, tachypneic, and febrile without apparent source of infection. At that point, he was begun on oral prednisone (PDR), 2 mg/kg/day. After 1 wk, he became afebrile with normal respirations. His adenopathy dissolved along with resolution of interstitial infiltrates. After 4 wks of PDR therapy, weight was 8.7 kg, serum immunoglobulins were normal, and spleen tip was just palpable. After tapering of PDR, all indicators of disease reverted toward pre-PDR values. Reinstitution of PDR again changed all parameters in a beneficial manner. We conclude that PDR may be effective in treatment of the AIDS prodrome.

Immune complexes in cerebrospinal fluid and serum of neurological patients: Possible intrathecal formation in bacterial meningitis and herpetic encephalitis

We assayed immune complexes (IC) by Particle Counting ImmunoAssay in the serum and cerebrospinal fluid (CSF) of patients with various neurological disorders. In pyogenic meningitis, the levels of IC sharply increased 4–8 days after onset with a fall before the 10th day of the disease. In herpetic encephalitis the IC and antibody levels started to increase about 12 days after onset. The IC persisted at high values for 3–4 weeks, whereas the high antibody titers persisted for several months during the follow-up. In these 2 groups of patients IC were probably locally produced as indicated by the lack of correlation with the IC levels in the serum. We did not detect any significant increase of IC in the serum and CSF of patients with multiple sclerosis (N = 48) or with acute idiopathic polyradiculoneuritis (N = 11). Using another technique based on the determination of IgG and C4 in polyethylene glycol precipitates we also failed to detect any significant increase of IC in multiple sclerosis sera.

Delta agent infection in Melbourne.

Evidence of infection with delta agent was sought in 284 patients with acute hepatitis B referred to Fairfield Hospital, Melbourne, between July 1, 1981, and June 30, 1982 and in 127 hepatitis B surface antigen (HBsAg) carriers. Acute and convalescent serum samples from the patients with acute hepatitis B and single or multiple sera from the HBsAg carriers were tested for delta antigen (delta ag) and/or antibody (anti-delta) by solid-phase radioimmunoassay (SPRIA). Evidence of infection was detected in 14.4% of patients with acute hepatitis B and 7.1% of chronic HBsAg carriers. Eighty-eight percent of those in whom delta agent infection was demonstrated were intravenous drug users.

Elevated antibody levels against measles virus P protein in sera of patients with multiple sclerosis.

Immune precipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and densitometry was used to estimate immunoglobulin G antibody levels against the H, P, and M proteins of measles virus in the sera of 24 patients with multiple sclerosis and 24 serologically matched controls. Of the 24 multiple sclerosis sera, 5 showed a statistically significant increase in antibody titer to the P protein as compared with the control sera. Antibody titers to the H and M proteins in multiple sclerosis and control sera were not significantly different.

Use of I region-restricted, antigen-specific T cell hybridomas to produce idiotypically specific anti-receptor antibodies.

Abstract Murine T cell hybridomas bearing receptors for antigen plus I region gene products were used as immunogens in mice in an effort to raise anti-receptor antisera. The antisera were assayed for anti-receptor activity by the ability to inhibit interleukin 2 production by the T cell hybridomas stimulated by antigen and I region expressing antigen-presenting cells. The T cell hybridomas used in these experiments were made by fusing antigen-specific, I region-restricted BALB/c T cell blasts to the AKR thymoma, BW5147. Three groups of mice were immunized with the T cell hybridomas: (BALB/c X AKR)F1 animals, syngeneic to the hybridoma; (BALB.B X aKR)F1 animals, differing from the hybridomas at H2; and (C.B20 X AKR)F1 animals, differing from the hybridomas at Igh. Mice were immunized multiple times and sera from individual animals were assayed for anti-receptor antibodies. In all groups, some mice produced anti-receptor antibodies by the criterion that they were inhibitory in the assay mentioned above. The frequency of mice producing these inhibitory antibodies varied considerably between groups, with the (BALB.B X AKR)F1 animals producing these antibodies most frequently, and the (BALB/c X AKR)F1 animals producing them least often. All inhibitory antisera were idiotypically specific; they inhibited the response of the immunizing T cell hybridomas, but not the responses of closely related hybridomas with different specificities. Moreover, when they could be absorbed, the inhibitory antibodies could only be absorbed by the immunizing hybridoma. It is hoped that these antisera, and B cell hybridomas prepared from the immunized animals, will be useful in the elucidation of the structure of the receptors for antigen plus I region products on T cells.

Mithramycin impairs the release of 45Ca from bone induced by prostaglandin E2 or multiple myeloma sera. Implications for a novel means of local tumor control.

Some tumors release factors able to activate host osteoclasts. Mithramycin at sub-tumoricidal doses inhibits the release of calcium mediated by osteoclasts. If invasion of bone by a cancer requires activation of these cells, their intermittent "blockade' might impede the development of metastases to bone or their local extension. Fetal rat bones prelabelled with 45Ca were cultured in the presence of 10(-7) M prostaglandin E2, sera from normal individuals, or from patients with multiple myeloma. Additional samples preincubated for 3 h with 1 microgram/ml of mithramycin, were washed before culture. Compared with controls, prostaglandin E2 stimulated the release of 45Ca by 28% (5 experiments) and mithramycin inhibited release by 15% (3 experiments). Preexposure to this cytotoxic antibiotic before culture with PGE2 reduced the augmented release. Sera from 4 patients with multiple myeloma were incubated with 45Ca-labelled bones, some pretreated with mithramycin. An additional 29% release of 45Ca (4 experiments) was prevented by mithramycin. These results are consistent with the hypothesis that augmented release of 45Ca due to stimulatory factors such as prostagladins or factors in sera from patients with multiple myeloma can be partially inhibited by pretreatment with mithramycin. Possibly, intermittent blockade of host osteoclasts can impair formation of metastases to bone by cancers dependent upon their activation for this event, or reduce the extent of local invasion by established metastases. Modifying the behavior of a cancer by altering the host-response to factors which it releases represents a potential alternative to cytotoxic chemotherapy.

Autolymphocytotoxic antibodies in patients on dialysis awaiting renal transplantation.

Two-hundred eighty sera from 78 dialysis patients were examined for autolymphocytotoxic antibodies (ALCAs). Thirty-five (12%) sera in 18 (23%) patients had ALCAs. Eight patients had ALCAs in multiple sera. Two patients had ALCAs against both T and B lymphocytes, 2 against T lymphocytes, and 14 against B lymphocytes. In those sera with ALCAs against both B and T lymphocytes, broad thermal reactivity was present. In sera with ALCAs against only B lymphocytes, the majority were reactive at 5 C. Sixteen (89%) patients with ALCAs had some form of glomerulonephritis; five had rapidly progressive glomerulonephritis (RPGN) and three had systemic lupus erythematosis (SLE). Nine of 10 sera with a positive crossmatch against B lymphocytes from a renal transplant donor did not have ALCAs against autologous B lymphocytes; three sera with ALCAs against B lymphocytes and two with ALCAs against T lymphocytes had negative crossmatches against transplant donor lymphocytes. Positive crossmatches against donor B and T lymphocytes attributable solely to ALCAs were therefore uncommon in our patients. Nevertheless, patients at risk to develop ALCA, including those with SLE and RPGN, should be identified and their sera tested for ALCAs. Once identified, studies can be carried out to distinguish a positive crossmatch attributable to ALCAs from that attributable to HLA alloantibody.

Abnormal glutamic acid metabolism in multiple sclerosis

We have found extensive amino acid abnormalities in multiple sclerosis sera. The most consistent abnormality is an elevation in serum glutamate, which is most striking during relapses. The increase in glutamate in the patients does not occur sharply during the onset of the relapse. Instead it appears to rise gradually within a month or two prior to the onset of the clinical relapse, to reach a peak during the relapse and then to slowly decline.

Disproportionate elevation of the immunoglobulin G1 concentration in cerebrospinal fluids of patients with multiple sclerosis

We determined immunoglobulin G (IgG) subclass concentrations and studied their distributions in the cerebrospinal fluids of patients suffering from multiple sclerosis, other inflammatory neurological diseases, and non-inflammatory diseases of the nervous system in comparison with a control group. In addition, the four subclass concentrations were measured in serum specimens of the multiple sclerosis and control groups. These data were correlated with the extent of local IgG synthesis in the subarachnoid spaces of the patients belonging to the different groups. We found a selective elevation of the IgG1 subclass in the cerebrospinal fluids of multiple sclerosis patients, and there was only a very small overlap of the IgG1 ranges of the multiple sclerosis and control groups. No major differences were detected between the IgG subclass distributions in different courses of multiple sclerosis nor between multiple sclerosis and control sera. The group with non-inflammatory diseases showed a uniform elevation of all four subclasses and a greater overlap with the normal range. This latter feature was combined with an elevated IgG1 concentration in the group with other inflammatory diseases. It is concluded that locally synthesized IgG in the cerebrospinal fluids of multiple sclerosis patients consists mainly of IgG1.

Measles virus-specific antibodies and immunoglobulin M antiglobulin in sera from multiple sclerosis and rheumatoid arthritis patients

When rheumatoid factor in rheumatoid arthritis and multiple sclerosis sera was titrated by the fluorescent antibody method on measles virus-infected cells, there was a marked and variable drop in titer on acetone-fixed cells as compared with unfixed cells. This was accounted for by the failure of measles virus hemolysin-inhibiting (HLI) antibody of the immunoglobulin G class to bind to acetone-fixed infected cells. It was shown by staining unfixed and acetone-fixed measles virus-infected cells that rheumatoid factor in most rheumatoid arthritis sera combined with measles virus-specific hemagglutinin-inhibiting and HLI antibodies, whereas rheumatoid factor in multiple sclerosis sera combined only with HLI antibody. Rheumatoid factor of similar specificity was also observed in normal sera and occasionally in rheumatoid arthritis sera. Both rheumatoid arthritis and multiple sclerosis sera showed almost identical increases in average titer above normal of measles virus-specific fluorescent staining immunoglobulin G and HLI antibodies.

Myelin-binding antibodies in vitro. Immunoperoxidase studies with experimental allergic encephalomyelitis, anti-galactocerebroside and multiple sclerosis sera

To determine whether in vitro demyelinating activity of blood sera correlates with the presence of myelin-binding immunoglobulin, we studied sera from 7 rabbits with allergic encephalomyelitis induced with white matter homogenate (EAE), two with encephalomyelitis induced with myelin basic protein (MBP-EAE), two sensitized with galactocerebroside, and from 4 patients with multiple sclerosis (MS). Living, myelinated, organotypic cultures of fetal mouse spinal cord were incubated with serum, fixed, treated with peroxidase-conjugated anti-rabbit or anti-human immunoglobulins, and incubated with diaminobenzidine. Immuno-stained myelin, coursing across the cultures, was found with EAE and anti-galactocerebroside sera, and these sera had strong demyelinating activity. In addition, anti-galactocerebroside serum sometimes immuno-stained cell membrane of cells with elaborate branching processes, possibly oligodendrocytes. No immuno-stained myelin was found with MBP-EAE, MS or control sera. Of these, only MS serum had demyelinating activity. These results indicate that the demyelinating activity of animal sera is associated with a myelin-binding immunoglobulin, which in the presence of complement probably mediates demyelination. On the other hand, the demyelinating activity of MS serum does not appear to be associated with a myelin-binding immunoglobulin. Thus there are differences between the demyelinating activities of MS and EAE sera.

Antibody to Escherichia coli enterotoxin in meat-packing workers.

Meat-packing plant employees exposed to raw animal products had serological evidence of higher infection rates with heat-labile toxin producing enterotoxigenic Escherichia coli (LT-EEC). In those employees with multiple sera available for study over a ten-year period, a drop in mean anti-LT-EEC titer was observed, suggesting altered ecology of or exposure to the organism during this time. Prospective studies need to be done to determine if meat-packing workers actually experience a greater incidence of LT-EEC-induced diarrheal disease.

The inhibitory effect of multiple sclerosis and control sera on certain myelin‐catabolizing enzymes

The inhibitory effect of multiple sclerosis and control sera on certain myelin‐catabolizing enzymes

Neural blocking activity of multiple sclerosis and EAE sera.

Neural blocking activity of multiple sclerosis and EAE sera.

Neuroelectric Blocking Factors in Multiple Sclerosis and Normal Human Sera

Serum samples from 13 multiple sclerosis (MS) patients and ten normal human serum samples were applied to cerebral neocortex cultures and evaluated for their ability to block evoked electric activity. A high proportion of sera positive for neuroelectric blocking factors was found in both groups, and there was no substantial difference between serum samples from MS patients and those from normal human volunteers. Some of the neuroelectric blocking factors were thermolabile, others were thermostable, and still others may have been complement-dependent. It is concluded that the ability to block evoked electric responses in tissue cultures is a nonspecific serum property, and that it is not specifically related to the pathogenesis of demyelinating disease.

Measurement of human high density lipoprotein apolipoprotein A-I in serum by radioimmunoassay

A sensitive and specific double antibody radioimmunoassay for the major apolipoprotein (apo A-I) of human serum high density lipoprotein (HDL) was developed. Initial studies indicated that direct measurements of apo A-I concentration in whole untreated sera or isolated high density lipoprotein fractions yielded variable results, which were lower than those obtained in the corresponding samples which had been subjected to delipidation. Subsequently, it was observed that heating diluted sera or HDL for 3 hr at 52 degrees C prior to assay resulted in maximal increases in apo A-I immunoreactivity to levels comparable to those found in the delipidated specimens. This simple procedure permitted multiple sera to be assayed efficiently with full recovery of apo A-I.

An anti-immunoglobulin specificity in human multiple myeloma sera

A previously unreported specificity of anti-immunoglobulin has been found in four of five IgG multiple myeloma sera known to contain anti-immunoglobulin activity. The antibody activity was of the IgM class and thus discreet from the “M” component. The antigen (termed DJ) was detectable in all normal sera examined. DJ appears to be an isotype of the IgG 1 class of immunoglobulins and is present on 14% of IgG 1 myeloma proteins tested without relation to Gm type. All DJ (+) IgG 1 molecules tested are of the κ light chain type. DJ is present on the Fc fragment of IgG and may depend on structural or conformational determinants in the hinge region for its expression.

Avian leukosis antibody response in individuals given chicken embryo derived vaccines.

Under natural conditions, avian leukosis viruses (ALV) produce in chickens a variety of malignant disorders, manifested as solid invasive tumors, lymphomatosis, myeloblastosis, etc. As these agents are ubiquitous in conventional chicken flocks and are transmitted vertically from hen to egg, virtually all vaccines derived from conventional eggs are contaminated with ALV (1—4). Experimentally, some strains of Rous sarcoma virus (RSV) have produced tumors in nonavian species (5), including nonhuman primates (6). Because of this, the question of the potential oncogenicity of ALV to humans arises. There have been isolated reports of serologic surveys of ALV antibody in man (2, 3). These previous studies were limited to the detection of neutralizing antibody to the Bryan strain of RSV, a test only sensitive to subgroup A avian leukosis viruses. We have, therefore, examined multiple sera from individuals inoculated with ALV-contaminated vaccines for the presence of antibody by employing the broadly reacting COFAL (7) and the highly sensitive indirect immunofluorescence (FAB) tests. The COFAL test is able to detect antibody to ALV of any subgroup. In the FAB test, subgroup A, B, and C antigens were employed to broaden the antigenic spectrum to cover the three largest ALV subgroups. Neutralization tests were performed on some sera. The results of these studies constitute this report. Materials and Methods. Vaccines. All ALV-contaminated vaccines were produced in chicken embryos derived from conventional flocks and were known to be contaminated with ALV. The ALV-free yellow fever vaccine (YFV) was prepared in ALV-free eggs from seed lots also free from ALV (4). Sera. Pre- and postimmunization serum samples were obtained from the following individuals inoculated with ALV-contaminated vaccines: 35 volunteers who received formalin-inactivated influenza vaccine in single or multiple doses; 15 inoculated with formalin-inactivated epidemic typhus vaccine; 15 who were given live attenuated measles vaccine; 15 who received conventional 17D YFV; and 15 who were given live smallpox vaccine.

EVALUATION OF MULTIPLE SCLEROSIS SERA AGAINST MEASLES ANTIGEN IN THE COMPLEMENT FIXATION TEST.

The examination of 28 spinal fluids of MS patients and 22 spinal fluids of control persons showed no differences in the content of complement fixing antibodies against measles virus. In contrast to spinal fluid findings, higher antibody titers could be demonstrated in the serum of 70 MS patients than in 548 control persons. These differences were of biostatistical significance.

An Improved Benzene Extracted Complement Fixing Antigen Applied to the Diagnosis of the Arthropod-Borne Virus Encephalitides

Summary and Conclusions An improved, lyophilized and benzene extracted mouse-brain complement fixing antigen for the diagnosis of the arthropod-borne virus encephalitides is described. The procedure employed in its preparation is shorter, simpler and less dangerous than the original described by DeBoer and Cox. The antigens are of higher titer and more specific, when used in the optimal dilution for sensitivity, than those prepared in our laboratory by technics previously described. Benzene extraction appears to remove the fractions usually responsible for anti-complementary and non-specific reactions. Wassermann positive sera and sera from certain other non-syphilitic, but febrile patients, do not fix complement in the presence of this antigen even when the sera are inactivated at only 60 C. 56 C was found to be inadequate for syphilitic sera. Antigens so prepared can be lyophilized a second time with no change in their antigenic activity and such lyophilized products can be used after rehydration without further centrifugation. This product after lyophilization the second time is suitable for commercial preparation and distribution. It appears to be very stable, at least at 5 C. When this antigen is used in dilutions lower than the “optimal,” certain immunological overlapping can be demonstrated between Eastern and Western equine encephalomyelitis viruses as well as with St. Louis and Japanese B encephalitis viruses. California virus reacts very slightly, one way only, to Japanese B and St. Louis immune guinea pig sera. No immunological relationship was observed between the equine viruses and the St. Louis-Japanese B complex or the California virus. Overlapping between related viruses can be entirely eliminated by determining the optimal range of activity of each antigen with its homologous immune guinea pig serum and using the highest dilution of antigen in this range. In this dilution the antigen has maximal sensitivity and specificity. This usually represents a 1 to 5 per cent concentration by weight, of the original mouse brain. This same dilution is usually that most suitable for tests on human sera. Antigens, although slightly infectious (105 or 106 fold loss of original infectivity) lose their remaining infectivity with no loss in their antigenicity, after a few weeks storage in the liquid state at 5 C. Results are presented of tests with several antigens on nine series of multiple blood sera from Western equine, St. Louis and Japanese B encephalitis patients. The development of an excellent amniotic sac (chick embryo) antigen for the equine viruses is described and discussed. The preparation and limitations of a Japanese B chick embryo antigen and failure to produce a similar antigen for the St. Louis virus are also presented. Both Western equine and Japanese B chick embryo antigens were tested extensively on human encephalitis sera. None of the whole embryo antigens were of practical value because many normal human sera reacted with normal embryo.