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Effect of canine immune macrophages and canine immune serum on the growth of Ehrlichia canis.

The effect of canine immune macrophages and canine immune serum (CIS) on the growth of Ehrlichia canis was studied in vitro. Treatment of E canis organisms with CIS before they were inoculated into immune macrophage cultures, coupled with pre- and postinoculation maintenance of cultures in CIS, suppressed macrophage infection rates to 1% or less. Immune macrophages collected from E canis-carrier dogs and maintained in canine normal serum were more resistant to the intracellular growth of E canis than were normal macrophages maintained in the same serum. Ehrlichiae introduced into normal macrophage cultures maintained in canine normal serum multiplied within the macrophages and destroyed them. Maximal immunity to intracellular growth of E canis, maximal destruction of this organism, or both, required the interaction of E canis with both humoral and cellular factors in proper sequence.

Animals↗

[Inhibition of the growth of Ehrlich transplantable carcinoma in mice by antichorionic immune serum].

The literature data suggest the promotion of tumor cell implantation by chorionic gonadotropin. This hypothesis stimulated a study on the effect of antichorionic serum on the growth of Ehrlich solid carcinoma. An immune serum to chorionic gonadotropin obtained by immunization of rabbits was injected into mice with Ehrlich carcinoma transplanted under the skin of the back. Intramuscular injections of 0.1 ml were given twice a day for 14-21 days. A 45% inhibition of tumor growth (p less than 0.05) was observed after injection of serum with a high titer of antibodies to chorionic gonadotropin (1:10,000). It was matched by a much lower inhibition (21%), following injection of a low-titer serum (1:100). These results may be used as a rationale for development of an anahormone of chorionic gonadotropin for active immunization against chorionic gonadotropin.

Animals↗

Cutaneous basophil hypersensitivity in contact-sensitized guinea pigs. I. Transfer with immune serum.

Cutaneous basophil hypersensitivity, an immune inflammatory reaction characterized by infiltrates of basophils and a delayed time-course, was studied in guinea pigs contact sensitized with oxazolone. Routine histological techniques, employing ordinary paraffin sections, were modified to study this reaction. When biopsies of contact lesions were processed by these methods dense infiltrates of basophils could be demonstrated. Animals sensitized with complete Freund's adjuvant emulsified with oxazolone-keyhole limpet hemocyanin conjugates also developed delayed-in-time responses to contact challenge with oxazolone but not to picryl chloride. These hapten-specific delayed-in-time reactions also contained substantial numbers of basophils. Transfer of serum from actively sensitized guinea pigs resulted in specific accumulation of basophils at challenge sites of recipients. Thus, in this experimental system, cutaneous basophil hypersensitivity was found to be a hapten-specific delayed-in-time reaction that could be transferred with immune serum.

Adjuvants, Immunologic↗

Immune serum against anti-DNA-8-methoxypsoralen photoadduct.

Specific immune serum against photo-damaged DNA with the photoadducts of 8-methoxypsoralen (8-MOP) was obtained. The antigenic determinant is a polynucleotide chain with a mono-photoadduct: a coumarin moiety of psoralen linked through a cyclobutane ring to the 5,6-bond of thymine. The specific rabbit antiserum was used in the immunofluorescence method, for the detection of photodamaged DNA in the kinetoplasts and nuclei of unicellular organisms Crithidia luciliae, in the nuclei of snap-frozen tissue of mammals, and in human blood lymphocytes. Detection by the immunofluorescence method was limited by psoralen availability in situ; the psoralen concentration should be in the range 0.05-0.2 microgram/cm2 on specimens submitted to a topical application or about 1 microgram/ml in cell suspension. The long-wave ultraviolet light (UV-A) exposure dose applied to the nuclei should exceed 1 J/cm2.

Animals↗

Immunoprophylaxis and immunotherapy of respiratory syncytial virus-infected mice with respiratory syncytial virus-specific immune serum.

The effects of passive respiratory syncytial virus (RSV) immune serum given as preinoculation prophylaxis, postinoculation prophylaxis, and as therapy on d 5 after inoculation were evaluated in an illness model of RSV infection in BALB/c mice. Pre- and postinoculation prophylaxis reduced RSV replication in lung after primary RSV infection and prevented illness. Day 5 treatment did not affect peak titer of RSV in lung but resulted in more rapid recovery from illness than seen in untreated mice. Prophylaxis prevented antibody responses and lymphocytic infiltrates in lung after primary RSV infection. Treatment caused diminished antibody and pathologic responses. Prophylaxis increased susceptibility to reinfection, although illness after rechallenge was mild. Mice treated therapeutically were less susceptible to reinfection than mice treated prophylactically, but they also experienced mild illness after rechallenge. Passive antibody prophylaxis and treatment of RSV infection are promising approaches to attenuating lower respiratory tract illness from primary RSV infection. The ability to measure illness endpoints and pathology make the BALB/c mouse model of RSV infection a useful system for the preclinical evaluation of immunoprophylactic and immunotherapeutic modalities.

Animals↗

The three-year incidence of non-B viral hepatitis morbidity in a controlled trial of pre-exposure immune serum globulin prophylaxis.

A trial of pre-exposure immune serum globulin (ISG) vs. the prevailing policy of post-exposure ISG for the prevention of viral hepatitis was undertaken among 23,447 male and female military recruits in Israel, an endemic area for hepatitis A virus infection. We are reporting on the three-year follow-up of the incidence of non-B hepatitis among 12,835 male recruits, half of whom had been allocated to receive ISG on their first day of service, while the other half served as a non-vaccinated ("regular care") control group. Over the three-year period the overall cumulative incidence in the intervention group was half that of the control group (3.6 per 1000 vs. 7.2 per 1000; p = 0.0037). Protection was virtually absolute for up to nine months' follow-up. ISG may have provided prolonged partial protection well in excess of six to nine months. Between 10 to 18 months, half as many cases occurred in the intervention group as in the controls. By 18 months the difference in incidence between the study groups had reached a plateau and was highly significant (p = 0.002). Between 19 and 36 months the case increment in the two groups was similar. The benefit of receiving pre-exposure ISG remained evident three years after inception of the trial in this indigenous military population living in an endemic area.

Clinical Trials as Topic↗

Antibody to the hepatitis B surface antigen in immune serum globulin.

A collection of 1,278 lots of immune serum globulin (ISG) prepared by 19 United States manufacturers between 1962 and 1974 were tested for the hepatitis B surface antigen (HBSAg) and antibody (anti-HBS). Ten lots (0.8%), all of which were produced between 1962 and 1965 by two different manufacturers, were weakly positive for HBSAg (by radioimmunoassay). Seven hundred and seven lots (55.3%) were positive for anti-HBS (by passive hemagglutination). In general, titers of anti- HBS in lots of ISG were low, and the prevalence of anti HBS positive lots varied considerably among different manufacturers. ISG prepared from placental material was more commonly positive for anti-HBS than was ISG prepared from plasma. There was a striking overall increase in prevalence and titer of anti-HBS in ISG lots prepared during 1973 and 1974. This probably reflects the effect of elimination of strongly HBSAg-positive plasma units with the onset of routine screening for HBSAg which began in 1972.

Antibodies↗

Immune serum increases arenavirus replication in monocytes.

The U937 monocytic cell line was used to determine whether antibodies could facilitate infection and replication of the arenaviruses, Pichinde virus (PV) and Lassa fever virus (LFV). When high dilutions of PV-immune serum were added to cultures simultaneously with PV inoculum, virus replication was dramatically (1000-fold) increased. Low dilutions of this antiserum neutralized the virus. LFV also replicated in U937 cells. The presence of LFV-specific immune serum in the growth medium increased the viral titre as much as 10,000-fold. Addition of heat-aggregated IgG partially inhibited antibody-mediated enhancement, probably by inhibiting the binding of immune complexes to the monocytic cells.

Arenaviridae↗

Hepatitis B immune serum globulin and standard gamma globulin in prevention of hepatitis B infection among hospital staff: a preliminary report.

In May 1973 a controlled double-blind clinical trail with prophylactic injects of hepatitis B immune serum globulin (antibody titer by passive hemagglutination 1:355,000) and standard gamma globulin (1:100) was started in Sahlgren's Hospital, Göteborg, Sweden. The annual attack rate of clinical hepatitis B in the three departments studied had been 5 to 8 per cent during recent years. A total of 118 members of the hospital staff were prophylactically treated while 125 staff members were unwilling to participate and received no prophylactic treatment. During the first 20 months of study nine cases of clinical hepatitis B with jaundice occurred within the untreated group (7.2 per cent) while three cases (2.5 per cent) were observed in prophylactically treated individuals. After decoding it was found that 60 individuals had received specific hepatitis B immune serum globulin while 58 had received standard gamma globulin. Two of the three clinical cases of hepatitis B occurred within the standard gamma globulin group. Both groups included two individuals with transient antigenemia only and the standard gamma globulin group also included four individuals with antibody seroconversion.

Antibody Formation↗

Passive transfer of immune serum reduces the L1/adult ratio of worms recovered from the intestines of Strongyloides stercoralis-infected gerbils.

In Strongyloides stercoralis-infected gerbils it had been observed previously that the ratio of first-stage larvae to adult worms decreases after 3 wk of infection. Serum obtained from gerbils after this decrease in fecundity, but before worms were expelled, was transferred passively to other infected gerbils during the peak of worm fecundity. The immune serum caused a significant decrease in the L1/adult ratio but had no effect on the number of uterine eggs, the length, or the intestinal and ovarian ultrastructure of adult worms. Apparently a factor(s) in the immune serum either killed eggs, after oviposition, or the hatched larvae, without damaging the adult worms.

Animals↗

Serotypes of Plasmodium falciparum defined by immune serum inhibition of in vitro growth.

In vitro growth inhibition assays were used to detect antigenic differences among geographically distinct strains of Plasmodium falciparum. Owl monkeys were immunized against the Camp and FCR-3/FMG strains of P. falciparum by infection, drug treatment, and rechallenge with homologous parasites. Camp-immune monkey serum was used to inhibit the in vitro growth of eight strains of P. falciparum. Inhibition was maximum for the homologous Camp strain (an average of 62% inhibition by 100 ml/litre Camp-immune serum). Four other strains were inhibited to a lesser degree, and three strains (FCR-3/FMG, FVO, and Smith) were not significantly inhibited by Camp-immune serum at concentrations as high as 400 ml/litre. FCR-3/FMG-immune serum at a concentration of 50 ml/litre caused significant inhibition of the FCR-3/FMG strain, but not the Camp strain. Thus Camp and FCR-3/FMG strains appear to bear distinct antigenic determinants recognized by the homologous, but not the heterologous, antiserum. Inhibition of in vitro growth by immune serum may be useful for serotyping P. falciparum and may have application in the selection of strains for inclusion in a malaria vaccine.

Animals↗

Failure of immune serum globulin to prevent hepatitis B virus infection in infants born to HBsAg-positive mothers.

Twelve infants, born to mothers with hepatitis B virus infection, were inoculated within 7 days of birth with immune serum globulin containing antibody to hepatitis B surface antigen (HBsAg) titers of 1:32 to 1:64 as measured by passive hemagglutination. Six of nine infants (66.7%) born to HBsAg-positive carrier mothers became HBsAg-positive within 3 mo of age. In addition, two of three treated infants born to mothers with acute hepatitis B during the delivery period also developed HBsAg. The hepatitis e antigen was detected in four of five carrier mothers and in two mothers with acute hepatitis, whose infants subsequently became HBsAg positive. In addition, hepatitis B-specific DNA polymerase activity was detected in the seven HBsAg-positive mothers who transmitted the virus to their infants. All eight infants have remained persistently HBsAg positive. Thus, the immune serum globulin containing low-titer antibody to HBsAg is not protective when given to infants born to HBsAg carrier mothers or to mothers with acute hepatitis B during the delivery period.

Carrier State↗

Prevention of hepatitis type B with specific immune serum globulin.

In order to evaluate the preventive value of specific immune serum globulin against hepatitis type B, we have used this immune globulin in required doses in 12 patients (10 with AU antigen negative and 2 with AU antigen positive) with chronic renal failure who required maintenance hemodialysis for a period of 15 months, and we were able to prevent hepatitis type B in our dialysis patients.

Adolescent↗

In vitro effects of antiprotozoan drugs and immune serum on Pneumocystis carinii.

The in vitro survival of Pneumocystis carinii isolated from the lungs of rats with glucocorticoid-induced pneumocystosis has been evaluated by quantitation of uptake of vital dyes by the organisms after exposure to a variety of drugs, immune serum, and medium enriched with macrophage lysosomal enzyme. On the basis of these tests, it appears that pentamidine and chloroquine are directly lethal to the organisms, but that suramin, trimethoprim, and sulfamethoxazole are not. Neither immune serum supplemented with complement and lysozyme nor the supernatant of phagocytosing macrophages appeared to affect the viability of the organisms. Inhibitors of glucose uptake and metabolism appeared to kill Pneumocystis, but neither cycloheximide nor iododeoxyuridine was effective.

Animals↗

A simplified method for the preparation of immune-serum globulin.

This report describes the recovery of highly purified immune-serum globulin (ISG) from the 42% ethanol precipitate fraction (42% P) generated during the first step of albumin purification. The method consists of three purification steps: (1) reprecipitation of the 42% P at 20% ethanol, pH 7.2; (2) solubilization of ISG at 17% ethanol, pH 5.2, and (3) isoelectric precipitation of ISG at 25% ethanol, pH 7.2. ISG prepared by this method was homogeneous when subjected to immunoelectrophoresis and polyacrylamide gel electrophoresis, and gave a yield equivalent to 4.7 g/l plasma, corresponding to a final recovery of over 48%.

Chemical Fractionation↗

Opsonic activity of human immune serum on in vitro phagocytosis of Plasmodium falciparum infected red blood cells by monocytes.

In vitro human monocytes from normal blood donors ingest red blood cells infected with Plasmodium falciparum more efficiently than normal red blood cells (NRBC). The phagocytic activity of human monocytes for infected red blood cells (IRBC) is greatly enhanced by the addition of immune sera obtained from individuals living in areas with endemic malaria. In contrast, the addition of sera obtained from individuals recovering from a first infection, or pooled normal sera, does not result in increased phagocytosis of IRBC. The phagocytosis enhancing activity of immune sera is associated with the IgG fraction and IgG depleted sera do not stimulate phagocytosis. Enhanced immune serum mediated phagocytosis occurs as a result of opsonization of IRBC. This was demonstrated by experiments in which monocytes or IRBC were preincubated with immune serum prior to the phagocytic assay. The opsonic activity could be absorbed by IRBC but not by NRBC. The opsonization of IRBC and subsequent phagocytosis were also dependent on the stage of development of the intracellular parasite. IRBC containing schizonts and trophozoites were preferentially phagocytosed as compared with ring forms. The role of malaria induced surface alterations and/or malaria surface antigens in the opsonization of IRBC by immune sera is discussed. These experiments suggest that phagocytosis of P. falciparum IRBC by monocytes may play a role in the immune elimination of malaria infection in humans.

Erythrocytes↗

Adoptive transfer of immunity to oral challenge with virulent salmonellae in innately susceptible BALB/c mice requires both immune serum and T cells.

The mechanisms of immunity to salmonellae conferred by immunization with live vaccines were studied by adoptive transfer using the mouse-virulent strain Salmonella typhimurium C5 and innately susceptible BALB/c (ltys) mice. This organism cannot establish a sublethal infection in naive BALB/c mice. Animals immunized 2 to 3 months earlier with the S. typhimurium SL3261 aroA live vaccine were used as donors of serum, spleen cells, and mesenteric lymph node cells for naive recipients which were challenged orally with the virulent C5 strain. Simultaneous transfer of both immune serum and immune cells was necessary for protection. Simultaneously depleting the donors of CD4+ and CD8+ T cells by administration of antisera in vivo prior to cell harvesting showed that T cells were necessary for protection. The results demonstrate that both antibody and T cells are required for recall of immunity to oral challenge with virulent salmonellae in innately susceptible mice and suggest that the ability to elicit opsonizing antibody in addition to cell-mediated immunity is important for optimal protection induced by salmonella vaccines.

Animals↗