bluuebunny/crossref_metadata_2025 huggingface dataset:
Tric Type K, a New Immunologic Type of Chlamydia Trachomatis
Authors: Kuo Cho-Chou , Wang San-Pin , Grayston J. Thomas , Alexander E. Russell
Published: December 2022

Twelve Chlamydia trachomatis strains have been isolated from cervical and urethral swabs in a venereal disease study in Seattle that are identical immunologically in the micro-immunofluorescence-(micro-IF) typing test. These strains are closely related immunologically with the LGV type-L3 but easily separable from the other 12 TRIC and LGV types. Cross-absorption removed micro-IF heterologous antibody between the new strains and L3. A series of studies of biologic characteristics (infection of cell culture, mouse brain, and monkey eye) showed clearly that the new strains were not LGV but were similar to other TRIC strains. It is proposed that a new immunologic type be established—TRIC type-K.
Enhancement of Immune Response Potential of Mouse Lymphoid Cells Fractionated over Insolubilized Conjugated Histamine Columns
Authors: Shearer G. M. , Weinstein Yacob , Melmon Kenneth L.
Published: December 2022

Spleen cells from (BALB/c C57BL/6)F1 mice which had been immunized 7 days earlier with SRBC were either fractionated over columns of histamine-rabbit serum albumin-Sepharose beads or were untreated. Equal numbers of filtered and unfiltered cells were injected into irradiated, syngeneic recipients with SRBC. The kinetics of direct and indirect PFC generated by the two groups of transferred cells were similar. However, 4.5 more direct and 5.8 more indirect PFC were detected per unit number of spleen cells injected in the recipients of the filtered than in those of the unfiltered cells. In addition to enhanced PFC responses, spleen cell fractionation over the histamine-coated beads also resulted in slight but significant increases in the numbers of nucleated cells per recipient spleen. These findings indicate that removal of cells adherent to histamine-coated beads enhances the humoral immune response potential of mouse spleen cells. A similar result was obtained when histamine column-fractionated thymocytes were injected together with unfractionated bone marrow cells. In contrast, no enhancing effect was observed when fractionated marrow cells were mixed with unseparated thymocytes. Fractionation of spleen cells over poly-L-lysine-Sepharose columns, which bind mouse spleen cells and are similar in charge to histamine-rabbit serum albumin-Sepharose, had no detectable immunologic effect on transferred spleen cells. The possibility that the columns of insolubilized conjugated histamine retain thymus-derived lymphocytes which express amine receptors and function to regulate humoral immunity is considered.
Spontaneous “Autoimmune Disease” in Nude Mice
Authors: Morse Herbert C. , Steinberg Alfred D. , Schur Peter H. , Reed Norman D.
Published: December 2022

Sera from 1- to 4-month-old congenitally athymic “nude” mice and their phenotypically normal littermates (LM) were tested for their capacity to bind double-stranded DNA and RNA. Sera from nudes bound significantly more DNA than did LM sera with no differences in RNA-binding capacity being demonstrable. Decline in the clinical state of nude mice was associated with increased serum DNA-binding capacity. Nude mice grafted with thymi at 1 month of age had reduced serum DNA-binding capacities when compared to non-grafted nudes. Immunofluorescent staining of kidneys from nude mice revealed the presence of significant amounts of IgM, IgG1, IgG2, and IgA immunoglobulins in the glomeruli of nudes but not LM mice. In some cases, immunoglobulin was associated with complement (β1c) deposition. The relevance of these findings to theories of autoimmunity is discussed.
In Vitro Studies on Allotype Suppression
Authors: Adler Louise T.
Published: December 2022

Spleen cells from heterozygous rabbits with b locus immunoglobulin allotypes b4 and b5 which had been exposed in utero to maternal antibodies against the paternal allotype (b4) were stimulated in vitro with solubilized T2 phage antigen. The cells of thoroughly suppressed rabbits formed T2 neutralizing antibodies which were identifiable only as having allotype b5. The presence of anti-b5 serum in the cultures inhibited b5 expression and resulted in the formation of antibodies having the previously suppressed allotype, b4. Normal rabbit serum of the same allotypic specificity as the anti-b5 serum (b4) had no effect on the allotype distribution of anti-T2 antibodies. Although partial inhibition of b5 production occurred when the anti-b5 serum was removed from cultures after 5 hr, no reversal of b4 suppression was observed. Possible mechanisms accounting for these results are discussed.
Interspecies Variability in Antibody-Dependent Cellular Cytotoxicity (ADCC)
Authors: Zighelboim Jacob , Gale Robert P.
Published: December 2022

Lymphoid effector cells and antisera from several species were evaluated in allogeneic or xenogeneic test systems for their ability to mediate the cytolysis of radiochromium-labeled target cells, antibody-dependent cellular cytotoxicity (ADCC). Interspecies variation was observed at both the effector cell and antibody level. Species-dependent variability in either the structure or function of the effector cell receptor, the structure or function of the antibody molecule binding site for the effector cell, or a combination of these factors may be reflected in interspecies variability in ADCC activity.
Effector Cell Activity in Antibody-Mediated Cell-Dependent Immune Lympholysis
Authors: Kovithavongs Thavisakdi , Hottman Werner C. , Dossetor John B.
Published: December 2022

Lymphocytes from 52 normal individuals with ages ranging from 11 months to 92 years were determined for their effector cell activity, the ability to lyse an antibody-coated lymphocyte target, in the antibody-mediated cell-dependent immune lympholysis (ABCIL) system. Prepubertal males and females, and females in their active sexual age, have significantly lower effector cell activity than sexually active males or postclimacteric individuals. These findings suggest that male and female hormones may have some influence on effector cell activity in normal individuals.
The Synthesis of Secretory Component by Lymphoid Tissues in Vitro
Authors: Lai A Fat Rudy F. M. , McClelland David B. L. , van Furth Ralph
Published: December 2022

The results obtained with an in vitro culture technique demonstrate that free secretory component (SC) is synthesized by lymphoid tissues such as bone marrow, lymph nodes, spleen, tonsils, and adenoids, but not by the thymus. In most of the SC-positive cultures, bound SC (IgA-SC) is also found. These findings mean that the IgA-SC (secretory IgA) present in small amounts in the serum does not derive solely from spillover of secretory IgA and/or SC formed in the mucous membranes and glandular tissues, but that at least a part is derived from the lymphoid tissues. Cultures of thoracic duct lymphocytes and peripheral blood leukocytes from normal and pathological conditions (monocytic and chronic lymphatic leukemia) did not show synthesis of SC. The cellular localization of SC synthesis could not be determined, but it is conceivable that a common cell present in mucous membranes as well as in glandular and lymphoid tissues is the producer of SC.
Relationship of Carcinoembryonic Antigen to Blood Substances a and i: Evidence That the Antigenic Sites Are on Different Molecules
Authors: Cooper Amiel G. , Brown Michael C. , Kirch Mary E. , Rule Allyn H.
Published: December 2022

Extracts of human colon carcinomas have been assayed for the presence of blood group substance A and i activities using an automated quantitative hemagglutination-inhibition system. Carcinoembryonic antigen (CEA) activity was simultaneously assayed by the zirconyl phosphate gel radioimmunoassay. Isoelectric focusing of a perchloric acid tumor extract containing all three antigen activities indicated a similar pK1 for the three antigens. However, A- or i-specific affinity gel adsorption experiments showed that only the corresponding blood group antigen activity was removed from tumor extracts. CEA activity remained essentially the same after adsorption by these gels. Significant blood group activity could not be demonstrated in preparations of highly purified CEA or in CEA from which N-acetylneuraminic acid and fucose had been removed. Conversely, CEA activity was absent from preparations of purified blood group antigens and could not be unmasked by removal of the immunodominant β-galactose of the i antigen with β-galactosidase. These data provide evidence that the antigenic sites for CEA and blood group A and i antigens are located on different molecules in colon carcinoma extracts.
Leukocyte Aggregation: An in Vitro Assay for Cell-Mediated Immunity
Authors: Tom Baldwin H. , Jakstys Milda M. , Kahan Barry D.
Published: December 2022

An in vitro leukocyte-aggregation assay was developed to assess cell-mediated immune reactivity by hosts against their allotransplants. The assay, which measures the adhesion of host leukocytes to target cells, is caused by a specific recognition of donor antigens by sensitized lymphocytes. The formation of aggregates may reflect the release of chemotactic and migration inhibition factors following recognition of specific target cell antigens, thereby attracting nonimmune bystander cells to the reaction foci. The test is simple to perform, brief, and requires small amounts of materials. Analysis of the patterns of reactivity of host cells suggests that the immunological specificity of the recognition event is consistent with the HL-A system in 80% of cases. Instances of disparities between predicted and observed reactions were attributed to lack of host sensitization, differences between serological and cellular HL-A cross-reactivities, and the potential participation of other histocompatibility systems.
Immunogenicity of Lowly and Highly Derivatized Dinitrophenyl-Bovine γ-Globulin in Different Strains of Guinea Pigs
Authors: Quijada Luis , Kim Young T. , Siskind Gregory W. , Ovary Zoltan
Published: December 2022

Guinea pigs immunized with lowly derivatized dinitrophenylated bovine gamma globulin produced large amounts of antibody directed against the carrier molecule and large amounts of antibody directed against the hapten. Guinea pigs immunized with a highly derivatized preparation did not produce antibody against the carrier but produced large amounts of antibody directed against the hapten. Both preparations provoked the production of about the same amount of anti-hapten antibody, but the lowly derivatized antigen provoked the production of anti-hapten antibody of higher average affinity.
Selective Impairment of Cell Antigenicity by Fixation
Authors: Gatti Richard A. , Östborn Anita , Fagraeus Astrid
Published: December 2022

The effects of fixation by formaldehyde, p-formaldehyde, and glutaraldehyde on H-2a and Moloney leukemia virus (MLV)-associated antigens were evaluated. Two cell types were employed: a) a MLV-induced lymphoma, YAC, which is passaged in ascites form in A/Sn mice or grown in long-term culture, and b) MLV-infected A/Sn and BALB/c mouse fibroblast monolayers. Antigenicity of the suspension cells was assessed by conventional absorbtions of antisera of predetermined titers to H-2a and MLV, and subsequent titration of these absorbed antisera by the 51Cr release cytotoxicity test against pertinent target cells. Antigenicity of fibroblast monolayers was assessed directly by a mixed hemadsorption technique. At low concentrations and short fixation periods, p-formaldehyde selectively impaired MLV antigens, preserving H-2a. At high concentrations and longer fixation periods, p-formaldehyde markedly reduced H-2a antigenicity while preserving MLV antigens. Fixation of either suspension cells or monolayers with glutaraldehyde (0.25% for 5 min at 20°C) markedly impaired both types of antigenicity.
Effects of Neoplasms on Inflammation: Depression of Macrophage Accumulation after Tumor Implantation
Authors: Snyderman Ralph , Pike Marilyn C. , Blaylock Barbara L. , Weinstein Phillip
Published: December 2022

The local accumulation of macrophages at sites of neoplasms may be a critical event in immunologically mediated tumor killing. Individuals with neoplasms, however, have been noted to have depressed monocyte chemotactic responsiveness in vitro. To determine the effect of neoplasms on macrophage migration, mice were implanted subcutaneously with either sarcoma or hepatoma cells and their macrophage migratory function quantified in vivo and in vitro. The ability of tumor-bearing animals to mobilize macrophages to an inflammatory site in vivo was depressed by as much as 61% by 6 days after tumor implantation. The in vitro chemotactic responsiveness of macrophages recovered from the peritoneal cavities of tumor-bearing animals was also markedly depressed. Macrophage migration was not affected by implantation of normal syngeneic or allogeneic tissues. In addition, the accumulation of polymorphonuclear leukocytes in vivo was not depressed in tumor-bearing animals. These findings suggest that neoplasms themselves may depress the host's ability to localize macrophages at inflammatory sites in vivo and thereby hinder immunologically mediated tumor destruction.
A Simple Method for the Determination of Complement Receptor-Bearing Mononuclear Cells
Authors: Gelfand Jeffrey A. , Fauci Anthony S. , Green Ira , Frank Michael M.
Published: December 2022

A simple method is described for the identification of mononuclear cells bearing the complement receptor. Gram-negative bacteria are directly fluoresceinated and then incubated in non-immune human serum, which leads to complement fixation via activation of the alernative complement pathway. Complement receptor-bearing mononuclear cells (CRMC) will form easily readable rosettes with these bacteria. The method requires no antibody preparation or purification, uses human serum rather than purified complement components or mouse serum, and gives results that correlate closely (r = 0.9015) with the EAC method of identifying CRMC. Furthermore, these indicator cells cannot bind to mononuclear cells via the E or IgG Fc receptors, eliminating this source of possible error. This method may be used for the identification of CRMC in suspension or frozen tissue sections.
Complexity of Factors in Sera of Different Mice That Affect MTV-Induced Mammary Tumor Cells
Authors: Blair Phyllis B. , Lane Mary-Anne , Mar Pearl
Published: December 2022

Specific spleen cell activity in microcytotoxicity assay can be altered by pretreatment of target mammary tumor virus (MTV)-induced mammary tumor cells with serum. Serum from both BALB/cfC3H females neonatally infected with MTV and BALB/c females horizontally exposed to MTV antigens will block specific spleen cell activity against isologous mammary tumor cells. On fractionation of sera, blocking factors are localized in the 7S fraction. The 19S fraction contains recruiting factors that are not detectable in the unfractionated serum; these factors are active against isologous tumors and are thus distinct from the tumor-specific recruiting factors previously described in the sera of tumor-bearing females, which are active only against the autologous tumor. Antibodies mediating complement-dependent cell lysis are also detectable after serum fractionation.
Resistance to Murine Leukemia in Mice Rejecting Syngeneic Somatic Hybrid Cells
Authors: Liang Weitze , Cohen Edward P.
Published: December 2022

(A/J × C3H/HeJ) F1 mice reject somatic cell hybrids of ASL-1 cells (A origin) and LM (TK)- cells (C3H origin), but die from leukemia within 10 days after the inoculation of approximately 106 viable ASL-1 cells. Mice rejecting hybrid cells survive for prolonged periods after challenge with otherwise lethal numbers of ASL-1 cells. The hybrid cells, rejected by syngeneic F1 recipients, retained their oncogenic potential as determined by the appearance and progressive growth of tumors in immunologically deficient nu/nu mice injected with the cells. Similar results were obtained using hybrids of a radiation-induced cell line (RADA-1), maintained by serial transfer in strain A mice and LM(TK)- cells. Syngeneic mice injected with RADA-1 × LM(TK)- cells failed to form tumors. Mice rejecting RADA-1 × LM(TK)- hybrid cells survived for prolonged periods after challenge with otherwise lethal numbers of RADA-1 cells.
Role of Mercaptoethanol and Endotoxin in Stimulating B Lymphocyte Colony Formation in Vitro
Authors: Metcalf Donald
Published: December 2022

Mercaptoethanol is necessary to permit B lymphocyte colony formation in semi-solid agar cultures of cells from normal mouse lymphoid organs. Transfer studies on developing colonies showed that, in part, this was a direct action on B lymphocyte colony cells but evidence was produced that in the presence of mercaptoethanol lymphoid organ cells release a factor promoting colony growth.

Endotoxin strongly potentiated B lymphocyte colony formation in vitro by a direct action on colony cells but in the absence of mercaptoethanol did not allow cell survival or proliferation.
Kinetics of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Jones Jeffrey M. , Amsbaugh Diana F. , Stashak Philip W. , Prescott Benjamin , Baker Phillip J. , Alling David W.
Published: December 2022

For the first 126 hr after immunization of mice with an optimally immunogenic dose (0.5 µg) of Type III pneumococcal polysaccharide (SSS-III), splenic antibody-forming PFC and serum antibody levels were measured at 2- and 8-hr intervals, respectively. PFC were detected at 28 hr after immunization and then increased through 86 hr after immunization; thereafter, the number of PFC remained nearly constant for the next 20 to 24 hr, and then began to decline. By contrast, serum antibody was first detected 60 hr after immunization. The accumulation of serum antibody continued to lag behined the increase in numbers of PFC by 16 to 20 hr until maximal serum antibody levels were attained; curves fitted to the values obtained for each parameter were nearly parallel. Further analysis of the data indicated that PFC increased in a stepwise fashion, producing a staircase pattern that would be expected for a synchronous proliferation of PFC with multiple recruitment events. Experiments in which mice were injected with a mitotic inhibitor at time intervals after immunization indicated that those precursor cells, capable of differentiating into PFC, were proliferating already by 12 to 24 hr after immunization. The size of the stepwise increases in the number of PFC decreased with time after immunization, indicating that recruitment from this pool of precursor cells decreased with time. By contrast, when PFC were measured in ALS-treated mice at 2-hr intervals after immunization, PFC were found to increase in a stepwise fashion; however, damping of the size of the stepwise increases did not occur and four additional rounds of recruitment and proliferation were observed after the first 86 hr after immunization. These findings allowed construction of a mathematical model to describe the effect of suppressor cells on the PFC response to an optimally immunogenic dose of antigen. In this model, the negative effect of suppressor T cells began 24 hr after immunization, increased progressively with time, and functioned to decrease the birth rate of PFC. This suggested the hypothesis that the negative effect of suppressor T cells was directed primarily toward B cell precursors of PFC, diminishing recruitment of PFC from this pool of cells by either halting proliferation of the precursor cells or preventing their differentiation into PFC.
Thymus-Derived Peripheral Lymphocytes from Human Newborns Inhibit Division of Their Mothers' Lymphocytes
Authors: Olding Lars B. , Oldstone Michael B. A.
Published: December 2022

Lymphocytes from human newborns inhibit division of their mothers' lymphocytes. Three days after we cultured equal numbers of cells from a mother and her baby in the presence of PHA, mitosis of the mother's lymphocytes was suppressed 13-fold compared to that of the baby's lymphocytes. At the end of 3 days the number of baby's lymphocytes were double those of the mother's. The survival rates and mean mitotic indexes of both pairs of cells were roughly equivalent (mean ± S.E: baby 2.4 ± 0.8; mother 2.6 ± 0.7), indicating that the lack of dividing lymphocytes from the mother was caused by inhibited division of the mother's lymphocytes, not enhanced growth of the newborn's cells.

The cell population in newborns that is responsible for the inhibition effect resides in the T cell-enriched population. Lymphocytes from one newborn were not able to inhibit division of lymphocytes from another newborn, suggesting that lymphocytes from newborns could continue to divide despite their inhibitory effect. Other experiments showed that actively dividing fetal fibroblasts, amnion cells from the newborn, and continuous T lymphoblastoid cell lines were unable to inhibit mitosis of lymphocytes of the mother.
Degranulation of Basophilic Leukocytes in Allergic Contact Dermatitis Reactions in Man
Authors: Dvorak Ann M. , Mihm Martin C. , Dvorak Harold F.
Published: December 2022

Basophilic leukocytes participating in allergic contact dermatitis in man underwent progressive degranulation over the course of several days after skin test. Less than 10% of basophils were degranulated in 24-hr reactions, but more than 60% were degranulated by 72 hr.

In the electron microscope individual basophil granules developed looser packing of their constituent particles, focal replacement of particles by finely granular material, and eventual loss of all visible granule contents, changes that were progressive between 2- and 3-day reactions. However, granules undergoing these changes retained their limiting membranes as well as their identity in the cytoplasm. Empty granules appeared as membrane-bound vacuoles having the size and shape of intact basophil granules. In contrast to the findings in a case of basophilic leukemia, communication of granules with each other was extremely rare and communication of granules with the plasma membrane was not observed. However, membranebound vesicles, the size of pinocytotic vesicles and larger, were abundant in these basophils. Many vesicles were observed free in the cytoplasm, but others were in communication with the cytoplasmic granules or with the plasma membrane. Vesicles often contained finely granular material of the type occurring in altered granules and rarely contained material morphologically similar to the particles of intact granules. These vesicles may provide a transport mechanism for piecemeal release of basophil granule contents, in addition or as an alternative to the commonly accepted exocytotic pathway of basophil degranulation.
Cellular Immunity in Pyelonephritis: Identification of Suppressor Cell Activity of Spleen Cells in Response to Concanavalin A and Inhibition of Lymphocyte-Mediated L Cell Cytotoxicity
Authors: Williams Terry W. , Friedlander Arthur M. , Lyons Joseph M. , Braude Abraham I.
Published: December 2022

A population of fast sedimenting spleen cells was identified as the cause of impaired in vitro response of spleen cells to concanavalin A (Con A) in acute pyelonephritis in rats. These fast-sedimenting cells responded to Con A by suppressing the DNA synthetic response of normal spleen cells to Con A. In addition, spleen cells from acute pyelonephritic rats were significantly less able to mediate in vitro cell destruction in xenogeneic aggressor lymphocyte:target L cell mixtures. On the basis of these findings, the hypothesis is proposed that a suppressor cell can protect the pyelonephritic kidney against immunologically mediated tissue damage.
Regulation of the Immune Response to Tumor Antigens
Authors: Fujimoto Shigeyoshi , Greene Mark I. , Sehon Alec H.
Published: December 2022

A/Jax mice were rendered immune to the syngeneic and transplantable methylcholanthrene-induced Sarcoma 1509a by the surgical removal of the tumor 7 days after implantation; subsequent injection of tumor cells resulted in their rejection within 15 days.

The i.v. transfer of 107 to 108 washed thymus or spleen cells of tumor-bearing animals (TBA) to immune animals significantly inhibited the rejection of the tumor; this suppressive effect was entirely abolished by the treatment of these lymphocytes with anti-θ serum or anti-thymocyte serum (ATS) and complement before adoptive transfer. On the other hand, an equal number of thymus or spleen cells of normal animals or of animals bearing an unrelated tumor had no suppressive effect.

Treatment of normal syngeneic animals with ATS after tumor cell inoculation or splenectomy of TBA resulted in the suppression of the tumor growth. The serum of TBA had no effect on tumor growth in immune syngeneic mice.

Together these results suggest that TBA possess immunosuppressor T cells regulating negatively their immune response to the tumor.
Analysis of the Humoral Immune Response to Influenza Virus in Vitro
Authors: Braciale T. J. , Gerhard W. , Klinman N. R.
Published: December 2022

The induction of in vitro primary and secondary humoral immune responses to influenza virus in murine splenic explant culture is described. Anti-influenza antibody synthesized in vitro was detected and quantitated by a radioimmunoassay which utilized influenza coupled to bromoacetylcellulose. Both in vitro primary and secondary responses could be stimulated over a large range of virus doses. In vitro secondary responses were maximal when spleen donors had been immunized by the parenteral route although secondary type responses could be demonstrated as well after primary immunization by pulmonary infection. In vitro stimulation with influenza virus was relatively insensitive to inhibition at high antigen doses. The results are discussed in terms of the response to other antigens in spleen fragment culture.
Antibody-Induced Ultrastructural Changes of Malarial Sporozoites
Authors: Cochrane Alan H. , Aikawa Masamichi , Jeng Miji , Nussenzweig Ruth S.
Published: December 2022

Immunization with irradiated sporozoites produces a considerable degree of protection against rodent, simian, and human malaria. This protection is in part antibody mediated. Antibodies neutralize sporozoites (SNA), i.e., abolish their infectivity, and cause, in vitro, the formation of a thread-like precipitate on the parasites (CSP reaction). The present study was undertaken to characterize the ultrastructural aspects of antibody-sporozoite interaction. Gradient concentrated sporozoites of Plasmodium berghei and Plasmodium cynomolgi were incubated with immune and normal sera. Transmission electron microscopy revealed the presence of a prominent, thick coat surrounding the outer membrane of the sporozoites incubated in immune serum. The inner structure of these parasites appeared to be relatively unaltered. The coat was absent or minimal on parasites incubated in normal serum. Incubation of immune serum-pretreated sporozoites with rabbit anti-mouse γ-globulin conjugated with hemocyanin demonstrated the participation of immunglobulin in the formation of this surface deposition. Coat formation occurred also on the surface of metabolically inactive, non-secreting parasites such as formalin-treated sporozoites and parasites kept on ice. Metabolically inactive sporozoites failed, however, to produce a positive CSP reaction. Scanning electron microscopy and negative staining indicated that the anterior end of the parasites was free of coat deposition and that the thread-like precipitate (CSP) was located at the posterior end of the sporozoites.
Blocking of Certain Antigenic Sites in the F(ab) Region by Combination of Univalent Fragments of Rh Antibodies with Red Cell Antigens
Authors: Kunkel H. G. , Joslin F. , Hurley J.
Published: December 2022

Evidence was obtained in the present study substantiating the fact that divalent Rh antibodies are bound to antigenic sites on the red cells through only one of the two combining sites. Univalent F(ab) fragments bound to the red cell lacked such free-combining sites and showed a blockage of various idiotypic-like antigens which were readily detected when divalent antibodies or divalent fragments were used. The univalent fragments contained the antigens because they completely inhibited the divalent system. These antigens were detected with idiotypic antisera made to a variety of monoclonal Ig through the use of certain very heterogeneous anti-Rh sera. As is the case with many other idiotypic systems, combination with antigen which blocked the antibody-combining site eliminated the reactivity.

Advantage was taken of these differences between divalent and univalent antibodies to determine antigens in the F(ab) portion of Rh antibodies which are or are not blocked by such combination with the red cell. A variety of subclass and genetic markers found in the constant regions of the F(ab) units were not affected. However, in addition to the idiotype-related antigens a series of V region markers, particularly those determining various V region subgroups, were not detected after combination.
Defining the Heterogeneity of Anti-Tumor Antibody Responses
Authors: Segal Gary P. , Klinman Norman R.
Published: December 2022

Studies were undertaken to analyze the humoral in vivo and in vitro antibody response of BALB/c mice to a syngeneic MSV-induced tumor cell line. With a sensitive radioimmunoassay, sera obtained from individual progress or and regressor mice were shown to vary greatly in total tumor-specific antibody concentration as well as immunoglobulin class distribution of the antibody, but no significant difference existed between the groups of progressor and regressor mice as a whole. In addition, serum antibodies from all animals were shown to have extensive cross-reactivity against a variety of cell lines chosen to share one or more antigens with the cell line used for immunization. Conversely, when in vitro fragment cultures of splenic tissue from progress or and regressor mice were stimulated with tumor-related antigen, differences in responsiveness among normal, progressor, and regressor mice were observed. In addition, antibodies derived from fragment cultures displayed several different cross-reactivity patterns all of which were more restricted in specificity than serum antibodies.
Evolution of the Lymphoid System
Authors: Etlinger Howard M. , Hodgins Harold O. , Chiller Jacques M.
Published: December 2022

Leukocytes from the various lymphoid tissues of rainbow trout (RBT) were tested for their capacity to respond to the lymphocyte mitogens concanavalin A (Con A), lipopolysaccharide (LPS), and purified protein derivative of tuberculin (PPD). Thymocytes responded to Con A but not to LPS or PPD. In contrast, leukocytes from anterior kidney were stimulated with LPS but not with Con A or PPD. Cells from spleen and peripheral blood were stimulated by each mitogen. However, the degree of stimulation at optimally stimulatory concentrations of each mitogen was distinctive. The finding that the patterns of mitogenic responses of cells from each tissue were significantly different suggested that there is lymphoid heterogeneity in the RBT with a unique tissue distribution.

The species source of serum utilized as a medium supplement appeared to be capable of markedly affecting mitogenesis. Thus, LPS and PPD stimulation occurred in medium supplemented with rainbow trout serum (RBTS). On the other hand, LPS and PPD stimulation was not observed in medium supplemented with fetal bovine serum (FBS), with the exception of peripheral blood leukocytes which were stimulated by LPS in culture medium supplemented with FBS. Con A stimulated leukocytes from each lymphoid tissue in medium supplemented with RBTS and, with the exception of cells from anterior kidney, also stimulated cells from each tissue in medium supplemented with FBS.

The kinetic profiles of the responses of peripheral blood leukocytes to Con A, LPS, and PPD suggested that the extent as well as the time required for maximal stimulation was dependent on the dose of mitogen.
Enzyme-Linked Immunoassay: Conjugation of the Fab′ Fragment of Rabbit IgG with β-D-Galactosidase from E. Coli and its Use for Immunoassay
Authors: Kato Kanefusa , Fukui Hideo , Hamaguchi Yoshitaka , Ishikawa Eiji
Published: December 2022

A method for the conjugation of the Fab′ fragment of rabbit IgG with β-D-galactosidase from Escherichia coli is described. The method consists of two main steps: treatment of the Fab′ fragments containing sulfhydryl groups with excess N,N′-o-phenylenedimaleimide, to introduce maleimide residues into the fragments, and then incubation of the dimaleimide-treated Fab′ fragments with β-D-galactosidase, which also contains sulfhydryl groups, to form the rabbit Fab′-β-D-galactosidase complex. More than 90% of the enzyme used can be converted to the Fab′-enzyme complex, and the complex is readily separated from free Fab′ fragments by chromatography on a Sepharose 6B column. The application of the rabbit Fab′-β-D-galactosidase complex for immunoassay of macromolecular antigens is shown by measuring human IgG by the sandwich method. The rabbit (anti-human IgG) IgG-coupled Sepharose 4B is incubated with human IgG and then with the rabbit (anti-human IgG) Fab′-enzyme complex, and the enzyme activity bound to the Sepharose is measured. In this way it is possible to determine as little as 0.3 fmoles of human IgG.
Specific Selection of Cytotoxic Effector Cells: The Generation of Cytotoxic T Cells in Rat Thoracic Duct Lymphocyte Populations Positively or Negatively Selected for Reactivity to Specific Strong Histocompatibility Alloantigens
Authors: Wilson Darcy B. , Marshak Ann , Razzino-Pierson Giuliana , Howard Jonathan C.
Published: December 2022

These studies consider the generation of cytotoxic T lymphocytes (CTL) from precursors (CTLP) present in rat thoracic duct lymphocytes after stimulation with strong alloantigens. Also, they explore the relationship between CTLP and “initiator” (I) lymphocytes responsible for specific GVH and MLI reactions.

Positively selected TDL populations prepared in bulk MLI cultures show enriched GVH and MLI reactivity for the selecting major histocompatibility complex (MHC) haplotype, but no cytotoxic activity, raising the possibility that I and CTLP may belong to different subpopulations, and the latter failed to differentiate or to survive under these culture conditions. Restimulation of these cells in Marbrook culture vessels with the original priming haplotype under conditions suitable for generating killer cells in vitro resulted in greatly increased specific CTL activity with accelerated kinetics soon after priming and normal kinetics later. These findings indicate that “memory” killer cells can be generated in a previously stimulated lymphocyte population that had no overt cytotoxic activity. Restimulation with third party haplotypes failed to give CTL activity either to specific or to third party targets.

Negatively selected TDL populations prepared by “filtration” through x-irradiated F1 rats, depleted of specific GVH and MLI responses, were also depleted of the ability to generate CTL in Marbrook cultures stimulated with the selecting haplotype. Stimulation with third party haplotypes, or with both third party and specific haplotypes together, gave CTL effective only against the third party target.
Structural Requirements for Arthritogenicity of Peptidoglycans from Staphylococcus Aureus and Lactobacillus Plantarum and Analogous Synthetic Compounds
Authors: Kohashi Osamu , Pearson Carl M. , Watanabe Yoshiro , Kotani Shozo , Koga Toshitaka
Published: December 2022

The comparative studies on the arthritogenicity of chemically well defined peptidoglycans (PG)2 from the cell walls of Staphylococcus aureus (FDA 209P) and Lactobacillus plantarum (ATCC 8014) showed that 1) a polymer of disaccharide-peptide with or without the presence of N-acetylglucosaminyl-ribitol-teichoic acid produced severe arthritis; 2) oligosaccharide-peptides with or without the special structure (N-acetylglucosaminyl-ribitol-teichoic acid in S. Aureus, a polymer of rhamnose and glucose in L. plantarum) produced severe disease; 3) disaccharide-heptapeptide-disaccharide with or without the presence of either glucose-ribitol-teichoic acid or a polymer of rhamnose and glucose appeared to be arthritogenic but much less effective for disease production; 4) N-acetylmuramyl-heptapeptide-N-acetylmuramic acid and disaccharide-hexapeptide were non-arthritogenic; 5) none of the synthetic N-acetylmuramyl-peptides, including tetrapeptide, produced the disease. Thus it is concluded that arthritis-inducing activity is related to the peptidoglycan moiety but not to the special structure, and the most important moiety responsible for disease production may be located in a chain length of two or more disaccharide units on PG subunits. However, it is discussed that non-arthritogenicity of peptidoglycans, including synthetic compounds, may be due to a lack of adjuvanticity in rats rather than a lack of antigenic determinant(s) responsible for production of arthritis.
Complement and Macrophage Activation
Authors: Bianco C. , Eden A. , Cohn Z. A.
Published: December 2022

Activated mouse macrophages, obtained after i.p. injection of thiglycollate medium, bacterial endotoxin, and many other agents, behave differently from the resident peritoneal cells: they are present in higher numbers, they spread faster on surfaces, they have higher pinocytic rates, and they secrete a number of neutral proteases which are not detected in non-activated cells. Activated macrophages ingest sheep erythrocytes prepared with IgM and complement (EIgMC) whereas non-activated macrophages only bind these sensitized erythrocytes.

Experiments aimed at identifying the several roles played by complement in the process of macrophage activation were performed both in vivo and in vitro.

The increase in cell number, the ability to spread, and the capacity to ingest EIgMC after the injection of thioglycollate medium and endotoxin were not observed in mice pretreated with cobra venom factor.

Fresh mouse serum incubated with inulin, zymosan or cobra venom factor, when added in vitro to recently explanted normal macrophages, induces marked changes of cell shape: increased membrane ruffling, appearance of microspikes and intense spreading which peaks after 1 hr at 37°C.
Interactions of Human C1̄ with C-Reactive Protein and Polyanion-Polycation Complexes
Authors: Claus D. R. , Siegel J. , Gewurz H.
Published: December 2022

C-reactive protein (CRP) is a potent activator of the classical C pathway when reacting with C-polysaccharide (CPS), choline phosphatides and certain polycations. Using a C1̄ binding assay we have shown that, like IgG, native CRP can bind purified human C1̄. This binding was increased approximately 10-fold when CRP was aggregated by heat or bis-diazotized benzidine treatment or when CRP was combined with its substrates including CPS and protamine sulfate. Human C1̄ was found to bind to and transfer from CPS-sensitized E pretreated with CRP (E-CPS-CRP) as well as with anti-CPS (rabbit) IgG, indicating that this interaction involved the C1̄ macromolecule, and suggesting participation of C1q; agglutination of E-CPS-CRP by purified C1q supported this conclusion. In agreement with earlier experiments performed in whole serum, interaction of polyanions (e.g., heparin) and polycations (e.g., protamine and poly-l-lysine) resulted in substantial C1̄ binding independent of the presence of immunoglobulins. This was quantitatively increased in the presence of CRP, and CRP markedly enhanced C consumption induced by interaction of these counterions in whole serum, suggesting that CRP may have an important role in C consumption by polyanion-polycation complexes.
Factor B on Lymphoid Cells
Authors: Halbwachs Lise , Lachmann P. J. , McConnell I.
Published: December 2022

When Cobra venom factor (CVF) is added to suspensions of well washed lymphoid cells, an enzyme is generated which will cleave C3 in normal human EDTA plasma.

The generation of this C3 convertase requires Mg++. Factor D enhances the reaction but is not necessary for it to occur. The effect is due to the anti-complementary factor in CVF and not to contaminating phospholipase A, as heating the CVF at 70°C abolishes activity. The lymphocytes are therefore supplying the activity of Factor B in the generation of the enzyme.

Normal peripheral blood lymphocytes, chronic lymphatic leukemic cells, and five lines of cells derived from Burkitt's lymphoma patients all show the Factor B activity; erythrocytes and thymus cells do not. The amount of C3-cleaving activity generated increases with cell number but at least 108 cells/ml EDTA plasma are needed to show an effect.

The C3 convertase formed by adding CVF to lymphoid cells elutes spontaneously from the cells.
Mesangiocapillary Glomerulonephritis: Some Correlations Between Ultrastructure and Complement Profiles
Authors: Swainson C. P. , Kay A. B.
Published: December 2022

Mesangiocapillary glomerulonephritis has characteristic structural features. Some patients have persistently low serum concentration of C3 with the presence of a C3 nephritic factor (C3 Nef). In the present report, 40 adult patients have been studied and the ultrastructural and immunofluorescent appearances of renal biopsies correlated with serum complement profiles.

On electron microscopy 22 patients showed “dense deposits” in the glomerular basement membrane and the remainder subendothelial deposits. Complement abnormalities were found in the serum of 10 out of 23 patients studied, 9 of whom had “dense deposit” disease. All had C3 Nef and low values of C3, and 5 had low values of C6 and C7. Two of these patients also had low C1q and C4 concentrations. C3 Nef was measured by crossed over immunoelectrophoresis and by disappearance of the B antigen on C3. There was a highly significant correlation between the two techniques.
Subunit Structure and Properties of Human C8
Authors: Kolb W. P. , Morgan W. T. , Müller-Eberhard H. J.
Published: December 2022

Studies of C8 subunit structure and conformation may provide important clues concerning the role of C8 in the formation of a transmembrane channel by the C5b-9 complex. Human C8, when examined by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis under reducing conditions, is composed of three polypeptide chains with the following molecular weights: α, 77,000; β, 63,000; and γ, 13,700. When using SDS and urea in the absence of reducing agents, two subunits are visualized: α-γ, 99,000 daltons, and β, 75,000 daltons. The α-chain appears to be largely concealed in the interior of the C8 molecule. This conclusion was deduced from the differential distribution of 125I after labeling of C8 by the chloramine-T method, either in its native form or under conditions which cause unfolding and dissociation into subunits. Whereas the β- and γ-chains take up 125I in proportion to their mass, the α-chain remains totally unlabeled in the native molecule.
Changes of the Complement Profile and Occurrence of Complement Breakdown Products After the Injection of Inulin or agg-IgG in Baboons
Authors: Lambert P.-H. , Houba V. , Perrin L. H. , Machnik F.
Published: December 2022

The interpretation of the complement profile in various clinical situations is often a difficult problem. In order to better define the changes occurring in vivo after the activation of the classical or of the alternative pathway, 13 baboons were injected either with aggregated human IgG (agg-IgG, 10 mg/kg) or with inulin (20 mg/kg) and plasma samples were collected serially. The following observations were made: 1) 4 hr after inulin or agg-IgG, C3 levels decrease to a minimum of 79 and 80% of the initial value, respectively. Native C3 antigen is decreasing faster than C antigen. 2) C1q remains constant in both experimental groups. C4 decreases to 84% 1 hr after Agg-IgG, then is normalized whereas, after inulin, it remains unchanged during 4 hr then strikingly increases to 188% after 24 hr. 3) Properdin decreases to 86% 1 hr after agg-IgG and to 79% 2 hr after inulin.
EAC14 and EAC4 Preparation with a Magnesium-binding Chelator, and SAC4 Inactivation by a Factor in Normal Serum
Authors: Linscott W. D.
Published: December 2022

Improvements have been made in the use of triethylenetetramine hexaacetic acid (TTHA), a strong chelator of Mg++ but not of Ca++, such that highly active EAC14 and EAC4 can be prepared by incubation of EA with TTHA-serum. It is essential to use a low ionic strenth, a long incubation period at 0°C, and to maintain a low pH (5.8) to avoid C3 contamination. The resulting cells are appreciably more active with limiting C2 than are cells prepared by the EAC1 plus EDTA-serum method, and stable to storage. IgG, IgM, and whole anti-Forssman antiserum all gave good preparations.

An unusual pH dependence was observed, in that SAC14 activity was generated between pH 5.6 and 7.0, and between pH 7.8 and 9.0, but not at pH 7.5. It was found that normal guinea pig serum contains a heat-stable factor which inactivates SAC4 at pH 7.5 in the presence of EDTA. This inactivation is such that there is a marked decrease in lysis with limiting C2, but no shortening of the Tmax.
Inhibition of Complement by a RNA Containing Extract Derived from Ehrlich Ascites Tumor Cells
Authors: Renk C. M. , Hoffman E. M.
Published: December 2022

Phosphate-buffered saline (PBS) extracts of whole Ehrlich ascites tumor cells (EATC) inhibit complement-mediated immune hemolysis. Attempts have been made to purify and characterize the active inhibitory factor in the tumor cell extracts. A combination of DEAE chromatography and phenol extraction yielded an inhibitory product that was approximately 80% RNA. Superior yields were obtained by direct phenol extraction of crude EATC extract followed by ethanol precipitation.

Phenol extracts of EATC material inhibited EAC1̄ and EAC1̄4. There was no effect on any of the other complement component intermediate complexes. EAC1̄ treated with the phenol extract were effectively inactivated in 15 min at 30°C. There was no inactivation of fluid phase C1̄ under the above conditions, but significant fluid phase C1̄ consumption was seen after longer incubation periods. In addition, phenol extracts of EATC precipitated with C1q after 2 days' incubation. Sucrose density gradient results indicated that the EATC RNA was heterogenous with respect to size, and it was more effective as an inhibitor than was RNA from other sources.
Studies on the Molecular Basis for C1̄s Action and Comparison of C1̄s with Other Tryptic Proteases
Authors: Roman D. P. , Andrews J. M. , Bing D. H. , Cory M.
Published: December 2022

One of the principal problems in current complement research is our understanding of the molecular basis of complement action. In order to probe the molecular biology and specificity of the serine protease, C1̄s, it was compared with three human proteases. The proteases examined were the serum enzymes C71̄s, thrombin and plasmin, and the pancreatic enzyme trypsin. An assay procedure using the simple synthetic chromophoric esters N-carbobenzoxy-l-tyrosine-p-nitrophenyl ester and N-carbobenzoxy-l-lysine-p-nitrophenyl ester as substrates was developed. A group of over 20 mono- and di-substituted benzamidines, known as competitive inhibitors of bovine tryptic enzymes, were used to probe the active sites.

The inhibition constants obtained were correlated with factors describing the lipophilicity and electronic character of the inhibitors using multiparameter regression analysis. Marked similarities were noted between C1̄s and the other enzymes. The difference between the enzymes with respect to their inhibition profiles can be interpreted on a molecular basis to account for some of the specificity shown by these enzymes.