bluuebunny/crossref_metadata_2025 huggingface dataset:
Properties of Rat Anaphylactic Antibody
Authors: Binaghi Ruben A. , Benacerraf Baruj , Bloch Kurt J. , Kourilsky Francois M.
Published: January 2023

The properties of the rat anaphylactic antibody have been investigated employing preparations containing both anaphylactic and nonanaphylactic purified anti-hapten (DNP) antibodies.

In zone electrophoresis, the anaphylactic antibody was found to migrate as a β-globulin, ahead of the main population of precipitating γ2-antibodies. Immunoelectrophoretic analysis did not reveal any precipitin line that could be ascribed to the anaphylactic antibody, probably because of its low concentration. Some heterogeneity was found in the γ-globulin region but a study of the passive hemagglutinating and lytic capacity of the antibody fractions with differing electrophoretic mobility failed to show any functional differences among these populations.

Ultracentrifugation studies have demonstrated that the anaphylactic antibody sedimented in a position intermediate between 7 S and 19 S components.

The anaphylactic activity was destroyed by heating at 56°C for 30 min.

These properties are discussed and compared with those shown by anaphylactic antibodies of other animal species.
Blood Group Antigens and Cross-Reacting Antibodies in Primates, Including Man
Authors: Wiener A. S. , Moor-Jankowski J. , Gordon E. B.
Published: January 2023

Orangutan red cells were typed for M blood factors with aid of cross-reacting anti-M reagents prepared from three different kinds of rabbit antisera, namely, against human type M red cells, against chimpanzee red cells, and against baboon red cells. The antisera had been rendered type-specific for human M red cells by absorption with human type N cells. Selective specificity for orangutan red cells was obtained by further absorptions with selected orangutan red cells. In this way, it was found that, among 24 orangutans, 12 were M positive and 12 were M negative. Absorption experiments indicate that human M cells probably have M factors lacking in orangutan M cells, whereas, conversely, orangutan M red cells probably have M factors lacking in human M cells. Although half of the orangutans tested lack M factors, none proved to have the NV factor. Thus, the M-N types of orangutans are not entirely comparable with the M-N types of man, just as had previously been found to be the case for chimpanzees. A suitable terminology to indicate these facts has been proposed.
Immunologic Studies of Pancreas-Specific Isoantigens
Authors: Metzgar Richard S.
Published: January 2023

Rhesus monkeys immunized with pooled Rhesus monkey pancreas extract and Freund's complete adjuvant developed pancreas-specific isoantibodies, and the electrophoretic mobility of the isoantigens is similar to that described in rabbits.

Passive transfer of antiserum or immune lymph node and spleen cells to recipients whose pancreas possessed the necessary isoantigens produced no histologic changes or functional alterations of the recipient's pancreas.

Autoimmunization of rabbits with their own pancreas incorporated in Freund's complete adjuvant failed to elicit auto- or isoantibodies and the pancreas of these immunized animals were normal grossly and histologically.

Immunofluorescent studies showed the rabbit pancreas isoantigens to be located in the cytoplasm of the apex of the acinar cells. The pancreas isoantigens of both rabbits and Rhesus monkeys were detected in pancreatic secretions.
Immunologic Comparison of Human Adult Thyroglobulin to Other Primate Adult and Human Fetal Thyroglobulins
Authors: Baur Sonia , Goodman Morris
Published: January 2023

Using human autoantibodies and rabbit and chicken heteroantibodies to adult human thyroglobulin, thyroid extracts from man, chimpanzees, old world monkeys, new world monkeys, pottos and nonprimate mammals were compared in Wilson modified Ouchterlony plates. Cross-reactions decreased as phylogenetic distance from man increased, with the decrease of cross-reaction being much greater for the human autoantibodies than for the rabbit or chicken antibodies on going from old world monkeys to more distant relatives. Thus, the less species-specific determinants on human thyroglobulin do not appear to be as autoantigenic as the more species-specific determinants. Adult human thyroid extract was also compared to 40 fetal thyroid extracts encompassing a broad range of gestational ages. Thyroglobulin was detected in most fetuses from 12 through 16½ weeks' gestation and in all fetuses after 16½ weeks. The quantity of thyroglobulin per milligram of extract protein was much smaller in early fetal thyroids than in late fetal and postnatal thyroids. Fetal thyroglobulin showed the same species specificity and antigenic specificity as adult thyroglobulin. A typical extract, whether from a fetal or postnatal gland, contained at least two types of saline soluble molecules with autoantigenic determinants. The more species-specific determinants, to which the majority of the autoantibodies were directed, were found on one type but not on the other. It is hypothesized that the less species-specific determinants predominate on a thyroglobulin fraction which more frequently gains access to the blood circulation than the thyroglobulin fraction with the more species-specific determinants. Thus a greater degree of immunologic tolerance or paralysis would develop to the less species-specific determinants than to the more species-specific determinants.
Facilitation of Tolerance in Weanling Mice by Enhancing the Proliferation of Donor Lymphoreticular Cells
Authors: Miller Joshua , Martinez Carlos , Good Robert A.
Published: January 2023

A latent period between injection of BALB/c spleen cells and grafting of BALB/c skin results in a substantially increased rate of graft acceptance by weanling DBA/2 recipients. Such a latent period does not facilitate tolerance induction when the donors are (C3H × A) F1 hybrids and recipients are weanlings of the C3H strain, a difference attributed to the lack of a stimulus to donor cell proliferation in this hybrid-to-parent combination. That stimuli to donor cell proliferation, other than the host antigens, also facilitate tolerance induction in weanling animals was also demonstrated. Five strong indifferent antigens were administered to donors 6 weeks before harvest of cells, and again to recipients following the cell transfer. A higher rate of tolerance induction was observed with BALB/c donors and weanling DBA/2 recipients, but the effect was much more evident with (C3H × A) F1 donors and weanling C3H recipients. Although females of the BALB/c and DBA/2 strains do not reject syngeneic male skin grafts, the graft-host interactions assay suggested that a histocompatibility difference associated with the Y chromosome does exist in these strains. Further support for such a difference was noted in the tolerance studies: When female cells were included in the BALB/c inoculum and the recipients were weanling DBA/2 males, the rate of tolerance induction was significantly increased over that observed when both donor and recipient were female. These studies suggest the following components in the mechanism of tolerance induction: a) proliferation of donor lymphoid cells which can be exaggerated by increasing graft vs host activity or by pretreatment with indifferent strong antigenic stimuli and b) a latent interval after cell transfer from donor to recipient and before attempted grafting during which it is proposed the donor cells proliferate in response to immune stimuli from the host. Observations presented here, and extended in the succeeding paper, suggest that it may be possible for immunologically competent cells to be committed to specific immune pathways to the exclusion of other pathways. The implications of these observations for transplantation theory and as a step toward development of immunologic tolerance in higher mammals are considered.
Studies of Rubella
Authors: Parkman P. D. , Buescher E. L. , Artenstein M. S. , McCown J. M. , Mundon F. K. , Druzd A. D.
Published: January 2023

Rubella virus interfered with propagation of enteroviruses, myxoviruses and arboviruses in GMK cultures. Mechanisms involved in production of this interference remain uncertain; no evidence of interferons has been found. Titers of infective rubella virus were measured most accurately by interference in GMK cultures incubated 10 days before challenge. These interference titers were not influenced significantly by specific type of challenge virus or its concentration. Rubella virus strains multiplied in 13 different cell systems including primary, continuous heteroploid and diploid cell lines. Tests in 11 of these showed similar rates of virus growth. Interference, however, developed differently in each culture system and could not be correlated with virus production. The virus was filterable through 300 but not 100 mµ Millipore filters. It was destroyed by heating, ethyl ether, chloroform and formalin. Thermal inactivation rates at 37°C were similar to those to rubeola, herpes simplex and influenza viruses. Propagation was not affected by IDU. No hemagglutinin or complement-fixing antigen has been shown. The findings support the original suggestion that rubella virus is unrelated to other known animal viruses.
Coccidioidin Skin Reactions
Authors: Wallraff Evelyn B. , Snow Isabelle B.
Published: January 2023

Two coccidioidins prepared by different methods were studied for skin test activity by comparison in guinea pigs, rabbits and human beings. There were 144 test comparisons in two series of guinea pigs, 22 in rabbits and 52 in humans.

A single intraperitoneal injection of the coccidioidins induced delayed hypersensitivity in guinea pigs. An enhanced hypersensitivity as measured by skin test response was obtained by incorporation of the coccidioidins in Freund's complete adjuvant.

Both coccidioidins were effective in eliciting skin responses in all species tested. When the test coccidioidins were diluted to have equal nitrogen content, their skin test reactivities in rabbits and human beings were the same.

A method for standardization of coccidioidins is suggested.
The Cell Cycle of Blast Cells Involved in Secondary Antibody Response
Authors: Sado Toshihiko , Makinodan T.
Published: January 2023

Spleen cells from primed donors were cultured with the test antigen in diffusion chambers. The cell cycle of blast cells was analyzed during the latent-log phase transition when the number of these cells remains constant. The estimated length of time for the various phases was: G2, 0.7 hr; M, 0.5 hr; G1, 0 to 1 hr; S, 5 to 6.8 hr; and the GT, 8 to 9 hr. No significant difference between the experimental and control groups was observed. The data on mean grain number per metaphase cell as a function of time after a single pulse exposure of these cells to H3-Tdr showed that the peak level occurred 3 to 4 hr after labeling, which corresponds to the latter third of the S phase. The significance of these results to the cytokinetics of antibody responses is discussed.
Some Factors Affecting the Secondary Immune Response in Tissue Cultures Containing Hydrocortisone
Authors: Halliday William J. , Garvey Justine S.
Published: January 2023

A complete secondary immune response with antibody production is possible in a defined culture medium, if primed rabbit lymph node fragments are challenged with antigen in vitro and incubated with hydrocortisone. These studies confirm the observations of Ambrose (7, 8).

Lymph node cell suspensions produced antibody only if prepared from tissue fragments which had been incubated in a hydrocortisone-containing medium for the preproduction phase (about 5 days). The hormone was not required for the antibody production phase of the cultures (tissue fragments or cell suspensions).

Unrelated antigens showed no cross-immunogenicity, although phytohemagglutinin stimulated a slight nonspecific response.

After priming injections in the feet, regional nodes were more responsive to challenge than were distant nodes. Spleen cultures were consistently negative under the conditions used.

Tracer studies showed that antigen was taken up by primed tissue in vitro, bound firmly and released only when antibody appeared in the culture fluids.
Antigen-Antibody Reactions in Allergic Human Tissues
Authors: Rappaport Ben Z.
Published: January 2023

In antigen-challenged nasal mucosa of seasonal hay fever patients, antigen, presumably bound to antibody, was identified in the cytoplasm of mucosa and gland epithelial cells, in histiocytes, pericytes, capillary endothelial cells and fibrocytes. In histamine-challenged tissue coated with antigen in vitro, fluorescence was demonstrable in surface epithelium, in gland cells, histiocytes and pericytes, but not in fibrocytes or endothelial cells. Fluorescent staining also suggests the posible presence of antigen and antibody in eosinophils of the allergic nasal mucosa.
Studies in Experimental Eosinophilia
Authors: Litt Mortimer
Published: January 2023

The draining lymph nodes of guinea pigs were examined histologically after the injection of BSA or hemocyanin into footpads. When a 10-mg dose of hemocyanin was administered, increased numbers of eosinophils were detected as early as 4 hr, and the peak response was seen at 12 hr. Margination, diapedesis and perivascular cuffing by eosinophils were prominent features in lymph nodes removed during the 1st hr, and such changes were evident as early as 5 min after the first exposure to hemocyanin. Similar findings were obtained with BSA, but the earliest changes were noted at 1 hr. When BSA, labeled with fluorescein, was injected into footpads, green fluorescence was noted in the draining lymph node as early as 5 sec. The experiments are consistent with the view that de novo synthesis of antibody may occur within 5 min of the first encounter of an organism with an antigen.
The Effect of Antigen on the Proliferation of Spleen Cell Suspensions from Tolerant Rabbits
Authors: Dutton Richard W.
Published: January 2023

Discussion and Summary

DNA synthesis is stimulated when homologous antigen is added to suspensions of spleen cells from previously immunized rabbits. This has been taken to represent the in vitro counterpart of the proliferative response seen in vivo and which leads to the appearance of differentiated cells which synthesize antibody.

The results described show that cells from a rabbit made tolerant by neonatal exposure to antigen do not respond to that antigen when challenged in vitro even after an intensive course of immunization. (It should be noted that small quantities of antigen were probably still present in the circulation at the time of sacrifice.) Thus, not only is no antibody synthesized but there is no proliferative response of precursor cells. In addition, it has been shown that simultaneous exposure of the cells to the antigen to which they are tolerant does not affect their response to a second antigen to which they are immune.

These findings are consonant with the reports of in vivo studies (1–4) referred to in the introduction and confirm the complete lack of a response at the cellular level in immunologically unresponsive animals.
Anti-Albumin Antibodies after Immunization with Whole and Enzyme-Digested Autologous Rabbit Albumins
Authors: Williams Ralph C.
Published: January 2023

Antibodies to rabbit (RSA), bovine (BSA), human (HSA) and horse (Ho.SA) serum albumins were produced by immunizing rabbits with various preparations of autologous albumins. The highest titers to HSA and Ho.SA were noted in animals immunized with relatively unaltered autologous albumin preparations; slightly higher titers against RSA were noted in animals given pepsin-digested autologous albumins. Antibodies to rabbit and human γ-globulin as measured by agglutination of incomplete antibody-coated cells were noted in some rabbits immunized with autologous albumin preparations. Antibodies were noted which showed specificity for sites uncovered by pepsin or rabbit spleen enzyme digestion of autologous albumins. Such antibodies were not inhibited or absorbed by unaltered autologous albumin.
Antibody Production by Determined Cells in Diffusion Chambers
Authors: Weiler Ivan Jeanne
Published: January 2023

Antibody production by immunologically determined cells has been followed in diffusion chambers. No antibody is produced by these cells in the first four days after tranfer to a new host. Then, although there is no new exposure to the specific antigen, antibody production commences and reaches a peak between days 8 and 15. At the same time, plasma cells begin to appear among clusters of lymphocytes within the chamber. A close association between mature plasma cells and fibroblasts could be demonstrated in 540 of 552 individually scored plasma cells.

Antibody titers at the time of maximum synthesis are higher within the chambers than in the host's serum or in empty chambers. This shows that the antibody is produced by the cells within the chambers, and not by the host cells.
The Immune Response of Rabbits Tolerant to One Protein Conjugate Following the Injection of Related Protein Conjugates
Authors: Weigle William O.
Published: January 2023

A high degree of tolerance was induced in rabbits by neonatal injection of picryl-bovine serum albumin (BSA). Twenty-seven of 36 rabbits were tolerant to both native BSA and picryl-BSA 3 months after birth. The other nine rabbits made an antibody response to both picryl-BSA and native BSA. The injection of the tolerant rabbits with picryl-horse γ-globulin (HoGG) resulted in a normal response to both the picryl group and the HoGG without interfering with tolerance to BSA. On the other hand, rabbits tolerant to picryl-BSA did not respond to picryl-RSA or DNP-BSA. The injection of the tolerant rabbits with sulfanil-BSA terminated the tolerant state to native BSA.

Data were given indicating that in normal rabbits the antibody response to picryl-RSA was directed to the hapten and a surrounding area on the conjugate.

Rabbits tolerant to picryl-BSA and injected with picryl-HoGG produced antibody that reacted with picryl-BSA but not with native BSA. When these rabbits were injected 1 month later with picryl-BSA, they again made antibody that reacted to picryl-BSA but not with native BSA.
Transfer of Immunity to Tetanus in Mice by Sensitized Homologous Lymph Node Cells
Authors: Nedelkoff B. , Heaton E.
Published: January 2023

Since Landsteiner and Chase first demonstrated passive transfer of sensitivity to picryl chloride by peritoneal exudate cells in guinea pigs (1), numerous studies have established the general nature of this phenomenon (2–5). In most cases biologic antigens, e.g., toxoids, have been employed by a variety of routes, and either circulating antibodies (2–4, 6, 7), delayed hypersensitivity (1) or both (8) have been detected.

The purpose of this study was to determine the extent of induced protection against lethal doses of tetanus toxin following passive transfer of sensitized lymph node cells (SLNC) in nonirradiated, adult, highly inbred mice. The mouse has been shown to be susceptible to tetanus toxin (9), and active immunization with tetanus toxoid has been produced in this animal (10).

Materials and Methods. The animals used were C3H/HeJ3 adult female mice. Fifty animals were immunized by injecting 0.25 ml of aluminum phosphate-adsorbed tetanus toxoid4 into the base of the tail and an equal amount subcutaneously into the interscapular region.
Immunochemical Aspects of the Cross-Reactivity between Groups A and C Streptococci as Detected by the Fluorescent Antibody Technique
Authors: Karakawa Walter W. , Krause Richard M. , Borman Earle K.
Published: January 2023

The cross-reactivity as detected by the fluorescent antibody method between Group A sera and Group C streptococci is dependent upon a rhamnose moiety that is shared by both Groups A and C carbohydrates and that, under certain circumstances, may be serologically reactive. The cross-reactivity is inhibited by A variant carbohydrate, and by a rhamnose disaccharide isolated from A variant carbohydrate by the rhamnosidase of McCarty. Cross-reactivity between Groups A and C carbohydrates is dependent, in part, upon the fact that this rhamnose disaccharide is a structural feature common to both antigens.
Agglutinating Factor in Myeloid Leukemia
Authors: Innis Michael D.
Published: January 2023

The sera of five consecutive patients with myeloid leukemia and the eluates from their leukocytes were examined for the presence of agglutinating factors by the technique of Gilboa-Garber and Nelken. The sera and eluates of five normal persons were similarly examined. Two patients with acute myeloid leukemia showed agglutinating factors in their sera; the eluates, on the other hand, of all five patients with leukemia showed the presence of these factors. In their avidity for cell surfaces they resemble warm incomplete anti-erythrocytic antibodies. None of the normal individuals examined showed any such abnormality. These factors appear to be either anti-leukocytic antibodies or some nonspecific agglutinating substance.
Prolonged Immunization
Authors: Lee C. L. , Davidsohn I. , Takahashi T.
Published: January 2023

Rabbit anti-mouse erythrocyte sera from each of 7 courses of immunization were studied with 9 immunologic techniques, namely, agglutinin titrations with antiglobulin serum, in serum-albumin medium, with papain treated cells, and in saline medium; hemolysin and complement fixation titrations; determination of combining ability with fluorescein and I125 labeled sera; and production of anemia in mice. Formalinized erythrocytes can be used successfully to study certain immunologic reactions. In vitro reactions, except for complement fixation and coating with fluorescent antibodies, did come near their peak levels in 2-course sera; after 4 or more courses, titers of all in vitro reactions were essentially parallel to the base line. Whether these tests are demonstrating different reactions of the same antibody or reactions of different antibodies requires further investigation. On the other hand, the anemia producing ability showed marked decrease in sera collected after more than 3 courses of immunization. Our studies permit the conclusion that no in vitro reaction has yet been found to serve as a reliable indicator of the in vivo potency of a rabbit anti-mouse erythrocyte serum.
Relationship of Passive Cutaneous Anaphylaxis to Antibodies in the Serum of Ragweed Hay Fever Patients after Injection Treatment
Authors: Connell John T. , Sherman William B.
Published: January 2023

Sera of treated hay fever patients, which contained blocking antibody in high titers (800 PNU or more), induced PCA in the skin of guinea pigs. There was a similar but less definite relation between high titers of hemagglutinating antibody and induction of PCA. Titers of skin sensitizing (Prausnitz-Kuestner) antibody have no relation to the activity of sera in PCA. Activity of the sera of treated hay fever patients in producing PCA was not affected by heating for 4 hr at 56°C or storage for 2 years at 4°C.
Further Observations on Cell Cultures Infected Concurrently with Mumps and Sindbis Viruses
Authors: Frothingham Thomas E.
Published: January 2023

The Sindbis virus plaque size enhancing property of mumps virus is shown to be closely associated with infectivity of mumps virus, and to be clearly different from mumps virus hemagglutinin and hemolysin. The enhancing property differs also from published descriptions of mumps virus cytolysin and mouse toxin.

Sindbis virus was recovered in much higher titer from doubly infected cultures which exhibited enhancement than from cultures showing normal Sindbis virus plaques. However, in chick embryo or human amnion cell cultures with liquid medium, Sindbis virus multiplication was similar whether or not concurrent mumps virus infection was present. The development of hemagglutinin in the mumps infected system was not affected by concurrent Sindbis virus infection.

Fluids harvested from doubly infected cultures interfered with vesicular stomatitis virus plaque development, while fluids from cultures infected solely with mumps or Sindbis virus failed to do so.

Heated allantoic fluid from influenza virusinfected eggs or fluid from rubella virus-infected cultures interfered with Sindbis virus activity in chick embryo or human amnion cell cultures respectively. Mumps virus infection inhibited this interfering activity.
Separation of Clostridium Perfringens Type D Toxins
Authors: Hauschild Andreas H. W.
Published: January 2023

A method was developed for the complete separation of Clostridium perfringens type D toxins.

ε and θ toxins were separated from each other and from the remaining toxins on a single column of diethylaminoethyl cellulose. After adsorption of θ toxin on sheep red blood cells, ε, α and κ toxins were separated in the same way.
Serologic Studies of Primary Epidemic Typhus and Recrudescent Typhus (Brill-Zinsser Disease)
Authors: Murray Edward S. , Gaon Jakob A. , O'Connor Jane M. , Mulahasanović Mujesira
Published: January 2023

Two distinct serologic patterns were observed in complement fixation (CF) tests on 172 sera from 70 patients suffering from infections caused by Rickettsia prowazeki.

The acute phase sera from 30 of the patients required 4 to 8 units of antigen in order to demonstrate maximum antibody titers; moreover, serum inactivation at 60°C for 30 min markedly lowered antibody titers. This group of patients also developed Weil Felix titers of 1:160 or greater between the 10th and 20th days of illness and showed no significant evidence of cross-reacting murine typhus CF antibodies. These 30 patients illustrate the clinical, epidemiologic and serologic characteristics of primary epidemic typhus. In contrast, acute phase sera from the remaining 40 patients gave almost identical antibody titers with both 1 and 8 units of antigen and these titers were not altered by serum inactivation at 60°C instead of 56°C for 30 min. This group of patients did not develop Weil Felix titers greater than 1:80 but did develop rising cross-reacting murine typhus CF antibodies. As a group these patients also demonstrated a dramatic rise in epidemic typhus CF antibody titers between the 7th and 10th days of illness. These 40 patients illustrate the clinical, epidemiologic and serologic characteristics of classic Brill-Zinsser disease. The sharply contrasting serologic patterns demonstrated in the immune responses of primary epidemic typhus and Brill-Zinsser disease add confirmation to Zinsser's hypothesis, which states that typhus occurs in a secondary (recrudescent) as well as a primary form. These findings also add to the accumulating data which point toward there being substantial physicochemical differences in the properties of primary and secondary immune response antibodies. The presence of the high antigen requirement (HAR) and heat labile (HL) phenomena can be demonstrated regularly in acute phase primary epidemic typhus sera; this observation implies that when diagnostic laboratories are testing for CF antibodies in such sera, 4 to 8 units of antigen and less than 58°C serum inactivation should be employed. The probable presence and significance of the HAR and HL phenomena in other infectious diseases is discussed.
Antigenic Properties of Human Mucous Membranes
Authors: Rose Noel R. , Bonstein Herbert S. , Mazow Jack B. , Arbesman Carl E. , Witebsky Ernest
Published: January 2023

Antisera specific for respiratory mucosa were produced by injecting rabbits with crude mucosal extract incorporated in Freund's adjuvant, and absorbing the resultant immune serum with kidney suspension. The antigenic properties of normal human respiratory mucosa, obtained from different levels of the respiratory tract, were studied by means of immunoelectrophoresis, double diffusion precipitation in agar, passive cutaneous anaphylaxis and in vitro anaphylaxis. No differences were found among the antigens of respiratory mucosa from different levels of the bronchial tree. There was cross-reaction between esophageal mucosa and respiratory mucosa, detectable by passive or in vitro anaphylaxis, but essentially none with mucosa of other levels of the alimentary tract. There was cross-reaction with lung, although the difficulties of obtaining pure lung preparations and pure preparations of segmental bronchus mucosa are pointed out. These reactions are apparently not due to blood group isoantigens or other group-specific antigens. These studies indicate the presence of tissue-specific antigens in respiratory mucosa.
Antiviral Activity of N-Ethylisatin β-Thiosemicarbazone in Vaccinia-Infected Mice
Authors: Pollikoff Ralph , Lieberman Melvin , Lem Nancy E. , Foley Edward J.
Published: January 2023

The anti-viral effect of N-ethylisatin β-thiosemicarbazone was determined in mice infected i.c. with vaccinia virus. The findings indicated that a single dose of 4 mg given as early as 72 hours before infection produced a 1 to 2 log lower PFU and LD50 titer and 80 to 100% net survival. When drug was given after virus inoculation, anti-viral efficacy was related to dose and time of treatment. In general, elementary or inclusion bodies were not observed in the brains of surviving drug-treated mice, and inflammatory changes were not suppressed. The absence of antibody or interferon in the brains of drug-treated mice prior to the 10th day following infection showed that protection was solely due to the virostatic action of N-ethylisatin β-thiosemicarbazone.
Differentiation of Lymphocytes Undergoing an Immune Response in Diffusion Chambers
Authors: Nettesheim Paul , Makinodan T.
Published: January 2023

No transformation of lymphocytes into histiocytes and fibroblasts was observed when cell-impermeable diffusion chambers containing rat thoracic duct lymphocytes were implanted intraperitoneally in unirradiated isologous or homologous recipients. When such chambers were implanted into irradiated heterologous recipients (mice), or when chambers containing these cells together with radiation-killed mouse cells were implanted into irradiated isologous recipients, the number of blast and immature cells increased with a T2 of approximately 13 hr between 2 and 6 days after culture. These cells were shown not to be of host origin. Pulse-labeling studies with tritiated thymidine indicated that they were derived mainly from deoxyribonucleic acid-synthesizing lymphocytes. No attempt was made to determine what cell type or types are the antigen-inducible progenitor cells, for previous studies indicate there is only 1 progenitor cell in approximately 106 lymphoid cells.
Observations on the Antibody Response of Rabbits to Mycobacterial Antigens
Authors: Daniel Thomas M.
Published: January 2023

The antibody response to soluble old tuberculin (OT), alum-precipitated purified protein derivative (PPD), OT incorporated in incomplete Freund's and whole killed mycobacteria in incomplete Freund's adjuvant has been studied in rabbits. Six of 12 animals injected with soluble OT and 3 of 8 given alum-precipitated PPD produced antibody measurable by hemagglutination techniques. This antibody was of short duration and was of the γ1M type exclusively. Five of 8 animals responded to OT in adjuvant and produced antibodies of both γ1M and 7 S-γ2 types. All animals responded to immunization with whole organisms in adjuvant. In these latter animals a transition from an early production of both classes of antibody, with γ1M antibody predominating in the earliest positive sera, to persistent exclusive synthesis of 7 S-γ2-immunoglobulin was observed. Following a second challenge an anamnestic response was not observed in any of the animals. That antibody which was synthesized following reinjection without adjuvant was of the γ1M type and was not accelerated in its appearance, suggesting a second primary rather than a secondary event. The data are in accord with the concept that the evocation of the 7 S-γ2 response requires a more intense stimulus than does the γ1M response. The production of 7 S-γ2 antibody, however, does not necessarily indicate preparation of the animal for a subsequent anamnestic response.
Purification and Some Properties of Anti-Testosterone Antibodies
Authors: Zimmering Paula E. , Beiser Sam M. , Erlanger Bernard F.
Published: January 2023

Antibodies to testosterone have been produced by immunization of ewes with conjugates of testosterone-17-hemisuccinate and bovine serum albumin suspended in Freund's complete adjuvant. Antibody has been separated from the sera by salt fractionation to obtain the globulin fraction, by dissolution of specific precipitates using a hapten analogue and by chromatography on DEAE-cellulose. The apparent specific precipitability of the antisera, and of all antibody-containing solutions derived from them, was markedly dependent on concentration. Our data can be expressed by the equation Aapp = A1 e-b(f-1) where Aapp is the apparent antibody titer at the dilution tested, A1 is the antibody titer of the undiluted test solution for which f = 1, f is the dilution factor, and b is a parameter which measures the degree of variation of precipitation of antibody with dilution. The concentration effect is probably not due to the solubility of antigen-antibody aggregates or to the changed configuration of proteins in dilute solution. It may be due to the presence of relatively weakly bound antibody molecules, or, more probably, to an increased possibility of intra-particulate interaction on dilution. The preferred method for specific purification of anti-testosterone antibodies was precipitation by heterologous antigen and dissolution of the precipitate by the hapten analogue, N,N-dimethylaminoethyl- 17β-(3-keto-4-androstenyl)-carbonate, followed by separation of the antibodies on Sephadex G-25. Two of the six ewes immunized produced antibodies which were bound by DEAE-cellulose, and which, when eluted, differed from unbound antibodies in that their specific precipitation by homologous antigen was inhibited by albumin or by some component associated with albumin.
The Isolation and Characterization of a New Component of Hemolytic Complement, C′3e
Authors: Inoue Kozo , Nelson Robert A.
Published: January 2023

Evidence is presented which describes the existence of an eighth fraction of normal guinea pig serum which is essential for the hemolytic action of complement (C′). This component, C′3e, reacts sequentially after C′3b and before C′3f. The intermediate complex, EAb C′1, 4, 2, 3c, 3b, 3e, has a half-life at 30°C of about 75 min. A method for the isolation from whole serum and for the measurement of C′3e reactivity is described. C′3e is depleted from fresh whole serum by antigen-Ab complexes. Purified C′3e appears to be depleted specifically by treatment with EAb C′1, 4, 2, 3c, 3b.

Initial studies indicate that C′3e is a macromolecule with a half-life at 56°C of about 41 min. As yet, there are no known co-factors or inhibitors of the hemolytic reactivity of C′3e.
Specific Suppression of Tumor Growth by Isolated Peritoneal Macrophages from Immunized Mice
Authors: Bennett Boyce
Published: January 2023

Methods were presented by which macrophages may be isolated from the peritoneal cell population of mice. These cells, and for comparison peritoneal lymphocytes and lymph node cells, were tested for immunologic activity by injecting a mixture of the test cells and tumor cells subcutaneously into irradiated mice. Each cell type, when obtained from immunized mice, was capable of suppressing the growth of the specific tumor cells. The results are discussed with respect to a possible specific immunologic function for macrophages in graft rejection.
Adsorption of Attenuated and Neurovirulent Poliovirus Strains to Central Nervous System Tissues of Primates
Authors: Harter Donald H. , Choppin Purnell W.
Published: January 2023

Attenuated and neurovirulent strains of all three types of poliovirus adsorbed to human and rhesus monkey brain and spinal cord gray matter at similar rates.

Combination with human brain homogenate inactivated the infectivity of attenuated and neurovirulent strains of the three poliovirus types; infectivity was restored by treatment with 6 M lithium chloride.

Results obtained suggest that the attenuated type 1 LSc, 2ab strain adsorbs to glass to a much greater extent than the neurovirulent Mahoney strain; this adsorption is markedly reduced by the presence of bovine albumin. The previously reported apparent failure of the attenuated LSc, 2ab strain to adsorb to primate neural tissues can be explained on the basis of extensive adsorption of this strain to glass in control tubes.

The results obtained do not support the hypothesis that attenuation of poliovirus reflects failure to adsorb to the cells of the primate nervous system.
The Separation and Physicochemical Properties of the C and D Antigens of Coxsackievirus
Authors: Frommhagen Laurence H.
Published: January 2023

The two antigens of coxsackievirus, Type B-5, separated by centrifugation in sucrose and cesium chloride density gradients, were shown to be very similar to the C and D antigens of poliomyelitis virus in their infectivity, complement-fixing properties, particle size and shape, and nucleic acid content. Certain practical aspects of the efficiency of separation of the C and D antigens in sucrose and cesium chloride density gradients were also discussed.
The Nature of Antibodies Synthesized during the Immune Response to Leptospira Biflexa
Authors: Hocker N. D. , Bauer D. C.
Published: January 2023

Agglutinating and hemolytic antibodies to Leptospira biflexa were examined after injection of rabbits with increasing concentrations of the spirochete. The magnitude of the agglutinating antibody response was dependent upon the antigen concentration. The response to the hemolytic antigen was not increased with increasing antigen but a persistent synthesis or renewed synthesis was observed which appeared to be quantitatively related to the level of antigen injected.

Secondary responses were initiated with a low dose of antigen. The agglutinating antibody response was similar to the primary response with that same dose of antigen. Hemolytic antibody synthesis, however, was increased more than twelvefold over the primary response.

This quantitative difference in the secondary response of the agglutinating and hemolytic antibodies might be explained on the basis of previous findings that anamnestic type responses are usually associated with the synthesis of low molecular weight antibody (7 S) but not heavy antibodies (19 S). However, examination of primary and secondary response antisera by density gradient ultracentrifugation and sensitivity to reduction indicate that both the agglutinating and hemolytic antibodies are rapidly sedimenting and sensitive to reduction throughout the immune response. Apparently anamnesis can be associated with macroglobulin antibody synthesis to certain antigens.
A Micromanipulator-Based System for Immunologic Analyses with Microliter and Submicroliter Reactant Volumes
Authors: Ringle David A. , Herndon Betty L.
Published: January 2023

An efficient, micromanipulator-based test system is described for the immunologic analysis of microliter and submicroliter volumes by complement fixation, hemagglutination and immunodiffusion in both agar and cellulose acetate. With this system it is possible to carry out all of these tests on a total antiserum volume of about 1.5 µl. Antigen volume requirements for these tests are correspondingly low. In regard to reactant concentrations the sensitivity of the micromethods compares favorably with the sensitivity of the corresponding macromethods.
Alteration of the Specificity of Lectin by Chemical Modification. II
Authors: Matsubara Shoji , Boyd William C.
Published: January 2023

The effects on the blood group specificity of lima bean anti-A of coupling to it diazotized amino derivatives of D-glucose and D-galactose were investigated. The galactose derivatives all enhanced the anti-B activity, p-aminophenyl-α-N-acetyl-D-galactosaminide (Ga III) being the most effective. Some of the glucose derivatives also enhanced the anti-B activity, but tended more to confer also an anti-O activity. The anti-B activity of these modified lectins was inhibited by 1 M d-galactose, and all activity inhibited by N - acetyl - d - galactosamine. Possible chemical mechanisms of the observed changes are discussed.
Studies on the Mechanism of the Cytolytic Effect of Sensitized Lymphocytes
Authors: Rosenau Werner , Moon Henry D.
Published: January 2023

Lymphocytes of mice sensitized by injection of homologous cells are capable of lysing, in tissue culture, cells of the same type used for sensitization. The serum of such animals also may contain cytotoxic humoral antibodies. The purpose of the present study was to ascertain if the cytolytic effect of the lymphocytes was due to their functional modification or to adsorbed humoral cytotoxic antibodies. The experiments were conducted to determine differences in the effects of ionizing radiation and hydrocortisone on the cytolytic effects of antiserum and lymphocytes from sensitized mice. The lytic effect of the sensitized lymphocytes was completely abolished by x-irradiation (15,000 r) whereas the cytolytic effect of antiserum was not demonstrably affected by the same level of irradiation. Furthermore, addition of hydrocortisone to homologous cytotoxic serum did not alter the lytic effect; previous experiments with hydrocortisone carried out under the same conditions had demonstrated a marked reduction in cytolysis mediated by sensitized homologous lymphoid cells. Since the cytolytic effect of sensitized lymphocytes was abolished by irradiation and was partially suppressed by hydrocortisone but the cytolytic effect of antiserum remained unchanged, it was concluded that lysis was related to functional modification of the lymphocytes and not to adsorbed cytotoxic antibodies.
The Preparation of Nucleotide-Protein Conjugates: Carbodiimides as Coupling Agents
Authors: Halloran Mark J. , Parker Charles W.
Published: January 2023

For some time it has been apparent that antibodies might be useful in the study of the fine structure of RNA and DNA. A major stumbling block has been the unavailability of a method which would render polynucleotides antigenic yet largely preserve their structural integrity. In approaching the problem of covalently conjugating polynucleotides to proteins as a means of stimulating antibody formation, we sought a procedure which would employ terminal nucleotide PO4 or OH groups for coupling. Two ways in which the terminal PO4 groups might be coupled covalently to protein would involve formation of a phosphodiester bond with protein seryl and threonyl residues or an N-P bond with protein ε-amino groups (Fig. 1, reactions 1 and 2 respectively). On the other hand, the OH group of a terminal sugar residue could react with protein carboxyl groups forming an ester (Fig. 1, reaction 3).
The Antigenic Structure of Treponema Pallidum, Nichols Strain
Authors: Miller James N. , De Bruijn J. H. , Bekker J. H. , Onvlee P. C.
Published: January 2023

Relatively tissue-free suspensions of Treponema pallidum, Nichols strain, were prepared by extracting the organism in rabbit serum-saline medium in equal parts. They were exposed to ultrasonic disintegration and the presence of antigens within the lysate and residue determined. Heat-labile and heat-stable (non-cardiolipin) antigens specific for T. pallidum, as well as antigens related to the lipopolysaccharide and protein (RP) components of the nonpathogenic Treponema reiteri, could be demonstrated. Further, the above antigens were shown to be unrelated to TPI antigen(s). The specific T. pallidum antigens within the lysate were found to be distinct from those within the residue.
Catabolism of γG-Globulin with Reduced Interchain Disulfide Bonds in Rabbits
Authors: Cohen George L. , Mannik Mart
Published: January 2023

The interchain disulfide bonds of rabbit γG-globulin were reduced and alkylated, but the H and L polypeptide chains remained assembled due to noncovalent interactions. The in vivo degradation of these altered molecules was compared to the degradation of unaltered molecules. The plasma half-lives were found to be identical in unaltered molecules, molecules with reduced interchain disulfide bonds and molecules with reoxidized bonds. Furthermore, molecules reassembled from separated H chains labeled with I125 and L chains labeled with I131 had plasma half-life comparable to the half-life of unaltered molecules. Therefore, the integrity of interchain disulfide bonds plays no role in the biological degradation of γG-globulin molecules.
Agar Gel Diffusion Method for the Study of Antigenic Components of Mumps Virus
Authors: Cuadrado Raul R.
Published: January 2023

A modified agar gel diffusion system for detecting mumps precipitating antibodies was investigated.

Mumps immune sera, when tested by a modified agar gel diffusion technique against concentrated soluble and viral antigens, showed a reaction only against the soluble antigen. When purified viral antigen was treated with peroxide-free ether for 15 to 30 min, a precipitating band, similar to that of the soluble antigen, was observed.

Agar gel diffusion methods may be used as a rapid tool for determination of precipitating antibodies to mumps virus, and possibly may be applied to the study of antigenically related parainfluenza viruses.
The Specificity of the Immune Response Following Injection of Bovine Serum Albumin into Neonatal Mice
Authors: Habicht Gail Sorem , Terres Geronimo
Published: January 2023

Neonatal mice were injected with 15 mg BSA and the temporal development of the immune response was followed. A biphasic response was observed. The possibility that the nodes in the immune response represented the production of antibodies of different specificity was investigated by using serologically cross reacting antigens (BSA and HSA). Mice injected at birth with 12.5 mg BSA showed no anaphylaxis or ADAR at the challenge or rechallenge with HSA. Mice injected at birth with BSA, challenged with BSA and rechallenged with HSA did show anaphylaxis and an elevated ADAR at the rechallenge. When mice injected with BSA at birth and challenged with HSA were rechallenged with BSA, the response to BSA was less than the response to BSA at the challenge. The specificity of the secondary response was discussed. When mice injected at birth with 12.5 mg BSA were challenged and rechallenged with HSA and SSA simultaneously it was found that the response to determinants on the SSA molecule developed before the response to determinants on the HSA molecule. Thus, the response to individual antigenic determinants on a single molecule does develop separately with time following neonatal exposure to the antigen.
The Antibody Responses of Rabbits and Rats to Hemocyanin
Authors: Dixon Frank J. , Jacot-Guillarmod Hubert , McConahey Patricia J.
Published: January 2023

The response of rabbits and rats to keyhole limpet hemocyanin (KLH) differed in a number of respects from responses to many other protein antigens. KLH is rapidly eliminated from the circulation and promptly degraded at least to the point of liberation of an I131 label in non-protein bound form. Maximum primary responses were elicited in rabbits by 2 mg of KLH and increases above this dose made little change in the response. Perhaps the most striking aspect of the antibody responses was that they consisted of considerable amounts of mercaptoethanol sensitive (MES) antibody. Primary responses of rabbits to KLH administered systemically were well sustained for several months and had approximately 50% MES antibody throughout the entire response. Six sequential responses of rats to systemically administered KLH were composed entirely of MES-19S antibody, and the secondary response was >30 times larger than the primary, indicating the operation of immunologic memory in this pure MES-19S antibody response. Even in responses to KLH in Freund's adjuvant or secondary systemic challenges, rabbits made significant amounts of MES antibody, and in few, if any, situations did the production of sizeable amounts of mercaptoethanol resistant antibody appear to interfere with the continued synthesis of MES antibody. Incorporation of KLH in incomplete Freund's adjuvant enhanced its immunogenicity approximately a thousandfold.