bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies of the Mechanism of the Human Platelet Release Reaction Induced by Immunologic Stimuli
Authors: Pfueller Sharron L. , Weber Susanne , Lüscher Ernst F.
Published: December 2022

Aggregated IgG coupled covalently with bis-diazobenzidine (BDB-IgG) and labeled with 3H-diazobenzene (3H-BDB-IgG) has been used to study the binding of soluble IgG aggregates to human platelets in relationship to the release of the contents of intracellular granules (e.g., serotonin). In washed cell suspensions a minimum of 0.14 to 0.2 µg 3H-BDB-IgG per 5 × 108 platelets (40 to 70 aggregates per cell) was required for the triggering of the release reaction and cell aggregation. Binding was independent of divalent cations. The Arrhenius plot gave a straight line between 0 to 37°C and a Q10 of 1.6. Neither inhibitors of the release reaction nor energy metabolism, nor formaldehyde fixation of the platelets affected binding. Bound 3H-BDB-IgG was not significantly eluted by IgG, bovine albumin (BSA), buffer, or plasma.

Binding to washed platelets was more strongly inhibited by human IgG than by F(ab′)2, bovine IgG, human albumin (HSA), or BSA. Plasma was an even more effective inhibitor of both binding and release. Plasma deficient in IgG or depleted of complement retained its inhibitory capacity. In the presence of plasma, at physiologic ratios of plasma and platelets, no release of serotonin was observed. Binding, although inhibited in rate, nevertheless occurred. It was enhanced by divalent cation chelation and had a Q10 of 2.5.

The release reaction of washed platelets to which 3H-BDB-IgG had been bound in the presence of HSA or BSA was also inhibited by the subsequent addition of plasma or plasma proteins (human IgG being more effective than bovine IgG, F(ab′)2, HSA, or BSA). 3H-BDB-IgG bound in the presence of either plasma or human IgG did not induce release when the platelets were subsequently suspended in media lacking these proteins. Thus, it appears that the platelet Fc receptor binds 3H-BDB-IgG by a process which is effectively inhibited by plasma, or by free IgG with an intact Fc, and to some extent by high concentrations of other proteins. The effects of bound IgG aggregates are dependent on the other proteins present both during binding and subsequently added. The mechanism of such receptor modulation and its implications in vivo are discussed.
Complement Activation by Measles Virus Cytotoxic Antibodies: Alternative Pathway C Activation by Hemagglutination-Inhibition Antibodies but Classical Activation by Hemolysin Antibodies
Authors: Ehrnst Anneka C.
Published: December 2022

Antibodies against measles virus hemagglutinating (HA) particles and hemolysin were shown to activate C differently. HA antibodies of rabbit or human origin activated C via the alternative pathway in cytotoxicity against chronically measles-infected cells. This cytotoxicity was expressed in C-4 deficient guinea-pig C or in rabbit C in the presence of 3 mM EGTA (ethylene-glycol-tetraacetic-acid) but not in 3 mM EDTA (ethylene-diamine-tetraacetic-acid). In contrast, human hemolysin antibodies activated C only via the classical way.

F (ab′)2 fragments from rabbit or human anti-HA IgG antibodies were as efficient in C activation via the alternative pathway as intact IgG antibodies with a corresponding hemagglutination-inhibition titer.
Antigen Recognition: The Specificity of an Isolated T Lymphocyte Population
Authors: Stashenko Philip P. , Schlossman Stuart F.
Published: December 2022

Guinea pig lymph node lymphocytes were separated into T and B cell fractions on immunoabsorbent columns. Separated cells were functionally distinct: T cells proliferated in response to ConA, PHA, soluble and alloantigen, whereas anti-Ig reagents only stimulated B cells. The in vitro proliferative response of guinea pig lymph node T lymphocytes was then shown to be highly discriminating when elicited by a series of structurally similar synthetic DNP-oligolysine antigens. Proliferation was always most extensive in response to the homologous, immunizing antigen, and less intense to cross-reacting DNP-oligolysines. Specificity of proliferation was maintained in the absence of both B lymphocytes and antibody secreting cells, suggesting that T cell recognition is not “acquired” from B cells or secreted antibody, but is a property inherent to the T cell.
Antigen-Specific T Cell Factors: A Fine Analysis of Specificity
Authors: Isac Ronit , Mozes Edna
Published: December 2022

The specificity of T cell factors produced in presence of synthetic polypeptide antigens was studied. Factors prepared with either one of the three antigens: poly(Tyr,Glu)-poly(DLAla)--poly(Lys), (T,G)-A--L, poly(Phe,Glu)-poly(DLAla)--poly(Lys), (Phe,G)-A--L, and poly(His,Glu)-poly(DLAla)--poly(Lys), (H,G)-A--L, successfully cooperated with B cells for antibody production to the homologous as well as to the other two immunogens. Furthermore, the activity of a (T,G)-A--L-specific factor was removed after passage through immunoadsorbents built of Sepharose coupled to: (T,G)-A--L, (Phe,G)-A--L and poly(Glu)-poly(DLAla)--poly(Lys), (G)-A--L, but not to poly(DLAla)--poly(LLys), A--L. No cross-reactivity was observed between (T,G)-A--L and poly(Tyr,Glu)-poly(Pro)--poly(Lys), (T,G)-Pro--L, at the level of T cell factors, as shown using the above approaches. These results lead to the conclusion that specificity of T cell factors, although not identical, is similar to that of antibodies.
Schistosomiasis in the Congenitally Athymic (Nude) Mouse
Authors: Phillips S. Michael , DiConza Joseph J. , Gold Jerome A. , Reid Willis A.
Published: December 2022

Athymic nude mice (Nu/Nu), heterozygous littermates (Nu/+), and thymus reconstituted homozygous animals (Nu/Nu TxR) were exposed to Schistosoma mansoni cercariae. A marked impairment in the development of peripheral blood eosinophilia and hepatic granuloma was observed in the Nu/Nu animal. In addition, host morbidity judged by organ weight, portal pressures and reticuloendothelial activity were less in the Nu/Nu animal. Thymus-reconstituted animals demonstrated eosinophilia, increased granuloma size, and morbidity equivalent to that shown by the Nu/+ heterozygous littermates. These studies indicate that the eosinophilia, granulomatous hypersensitivity, and ultimate host morbidity, associated with murine schistosomiasis, is dependent on thymic dependent lymphocyte function.
Enhancement by Irradiated T Cells of Human Plasma Cell Production: Dissection of Helper and Suppressor Functions in Vitro
Authors: Siegal Frederick P. , Siegal Marta
Published: December 2022

The addition of irradiated T-enriched lymphocytes to B cell-enriched fractions of human peripheral blood or to unseparated mononuclear cells stimulated the differentiation of plasmacytoid cells in culture with pokeweek mitogen beyond the synergy obtained by the addition of unirradiated T cells. This stimulation was observed in both the proportion and absolute number of plasmacytoid cells recovered from the cultures, and in the amount of IgM detected in culture supernatants by a hemagglutination inhibition assay. Irradiation-induced enhancement was observed with normal and hypogammaglobulinemic T cells, but not with monoclonal T cells from two patients. Inactivation of T cells by heating or by repeated freezing and thawing did not produce the same effects as did irradiation. These data suggest that cell-mediated suppressor function in man is selectively radiosensitive, while helper activity is not. Irradiation may be a useful method for the functional isolation of helper cells and for the manipulation of the balance between suppressor and helper cell activities.
Complement-Induced Histamine Release from Human Basophils
Authors: Hook William A. , Siraganian Reuben P.
Published: December 2022

Human serum activated with zymosan generates a factor (C5a) that releases histamine from autologous basophils. Previously we have presented evidence that this mechanism for C5a-induced release differs from IgE-mediated reactions. The effect of several pharmacologic agents known to alter IgE-mediated release was studied to determine whether they have a similar action on serum-induced release. Deuterium oxide (D2O), which enhances allergic release, inhibited in a concentration-dependent fashion the serum-induced reaction at incubation temperatures of 25 and 32°C. The colchicine-induced inhibition was not reversed by D2O. Cytochalasin B, which gives a variable enhancement of IgE-mediated release, had a marked enhancing effect on the serum-induced reaction in all subjects tested. The following agents known to inhibit the IgE-mediated reaction also inhibited serum-induced release at 25°C: colchicine, dibutyryl cyclic AMP, aminophylline, isoproterenol, cholera toxin, chlorphenesin, diethylcarbamazine, and 2-deoxy-D-glucose. These results suggest that the serum-induced release is modulated by intracellular cyclic AMP, requires energy, and is enhanced by the disruption of microfilaments. The lack of an effect by D2O would suggest that microtubular stabilization is not required. The data can be interpreted to indicate that IgE- and C5a-mediated reactions diverge at a late stage in the histamine release pathway.
Characterization of DNA Used to Assay Sera for Anti-DNA Antibodies; Determination of the Specificities of Anti-DNA Antibodies in SLE and Non-SLE Rheumatic Disease States
Authors: Locker Joseph D. , Medof M. Edward , Bennett Robert M. , Sukhupunyaraksa Surintra
Published: December 2022

Commercial 14C-labeled KB cell DNA, widely used to assay sera for anti-DNA antibodies, was chromatographed on benzoylated-naphthoylated-DEAE-cellulose (BNDC) and on hydroxyapatite (HAP). On BNDC, only 25% of the 14C label eluted with 1 M NaCl (KB fraction I) characteristic of ds-DNA. Fifty-five percent of the label eluted with 50% formamide-1 M NaCl (KB fraction II) characteristic of ss or denatured DNA. On HAP, however, none of the 14C label eluted with 0.2 M phosphate buffer as anticipated for ss-DNA, but, rather, all of the 14C label eluted with 0.4 M phosphate, characteristic of ds-DNA. After pretreatment with S1 endonuclease of Aspergillus oryzae, which selectively digests ss regions, however, 42% of the 14C label was lost from the 0.4 M phosphate peak. These results indicated that more than half of this 14C-KB-cell DNA preparation was ds-DNA with ss regions which was undetectable by HAP chromatography. 3H-ds-DNA and circular 3H-ss-DNA prepared from T7 and øX174 bacteriophage, respectively, were found to be chromatographically pure on both BNDC and HAP.

None of 10 non-SLE sera (rheumatoid arthritis 3, mixed connective tissue disease 4, scleroderma 1, ulcerative colitis 1, and pulmonary fibrosis with chronic active hepatitis 1), previously believed to contain anti-ds-DNA antibodies on the basis of KB cell DNA testing had detectable antibodies against KB fraction I or T7 DNA: all of 10 KB cell DNA positive SLE sera had antibodies against both. Additionally, none of the 10 non-SLE sera had antibodies against KB cell DNA when retested with DNA that had been pretreated with S1 endonuclease. Seven of these 10, however, as well as all 10 SLE sera, had antibodies against øX174 DNA, KB fraction II DNA and alkali-denatured T7 DNA.

The data support the conclusions that 1) false positive tests for anti-ds-DNA antibodies can result from contamination of ds-DNA with ds-DNA having ss regions, and 2) non-SLE sera do not contain antibodies specific for ds-DNA at levels comparable to those found in SLE sera but rather contain high levels of antibodies reacting with ss regions or mixed DNA.
On the Biochemical Racial-Index of the Ainu, Gilyak and Orokko in Karafuto (Saghalien)
Authors: Kishi Takayoshi
Published: December 2022

It is well known that we can classify all human blood into four different groups by isohemagglutination (Landsteiner (1), Jansky (2), Moss (3), v. Dungern (4) and Hirschfeld); namely,

Group I or O. The serum agglutinates the cells of groups II, III, and IV. The cells are agglutinated by no serum.

Group II or A. The serum agglutinates the cells of groups III and IV. The cells are agglutinated by the serum of groups I and III.

Group III or B. The serum agglutinates the cells of groups II and IV. The cells are agglutinated by the serum of groups I and II.

Group IV or AB. The serum agglutinates no cells. The cells are agglutinated by the serums of groups I, II, and III.

To explain this phenomenon, we must assume the existence of two agglutinable substances, A and B in the corpuscles and two agglutinins, α and β in the serum, distributed in the groups as follows.
Immunological Studies in Tuberculosis
Authors: Petroff S. A. , Stewart F. W.
Published: December 2022

Five series of experiments with guinea pigs sensitized with dead tubercle bacilli are reported, in which tuberculin hypersensitiveness was induced with dead tubercle bacilli. During the allergic state inoculations of living tubercle bacilli were made, with the use of approximately the same number of normal control animals. The sensitized animals outlived the controls an average of 46 days. The amount of macroscopic disease at autopsy in the former was much less extensive than in the latter, confirming the belief that dead tubercle bacilli can induce some degree of protection. A small series of guinea pigs was sensitized with Dreyer's defatted tubercle bacilli. We produced only a mild skin hypersensitiveness and even that not uniformly. No appreciable resistance could be observed in these animals. The average duration of life was approximately the same as that of the control animals.
Genetic Differences in BCG-Induced Resistance to Schistosoma Mansoni Are not Controlled by Genes within the Major Histocompatibility Complex of the Mouse
Authors: Civil Richard H. , Mahmoud Adel A. F.
Published: December 2022

Induction of nonspecific resistance to Schistosoma mansoni infection after the i.v. injection of viable BCG was investigated in outbred mice and a panel of inbred and H-2 congenic strains. Significant protection was induced in CF1, A/J, C57BL/6, C57BL/10, DBA/2, C57BR, and SJL mice. BALB/c mice were not protected whereas CBA and C3H mice expressed intermediate degrees of protection. Expression of the protective phenomenon is not controlled by genes within the MHC as shown by the marked differences in response between BALB/c and DBA/2 (H-2d) as well as between C57BR and C3H (H-2k) mice. H-2 congenic strains with C57BL/10 background (B10.A and B10.D2) were high responders. BALB.B10 mice carrying the high responder (B10) MHC on the nonresponder (BALB/c) background were not protected. The degree of splenic hypertrophy did not correlate with the expression of nonspecific resistance. These results demonstrate that, in addition to controlling specific immune responses, genetic differences influence the nonspecific protective phenomena related to BCG administration as well.
Immune Response to Immunization via the Anterior Chamber of the Eye
Authors: Kaplan Henry J. , Streilein J. Wayne
Published: December 2022

Exposure to alloantigen via the anterior chamber of the eye elicits a transient suppression of cellular immunity, whereas humoral immunity is preserved—i.e. F1 LI-ID. The majority of lymphoid cells inoculated into the anterior chamber are retained within the posterior segment of the eye. The latter serves as a depot of alloantigen, allowing the chronic egress of small numbers of cells into the vascular tree. The persistence of this antigen depot is essential to the development of F1 LI-ID. Since there is a preferential distribution of cells that migrate from the eye to the spleen, the functional integrity of the latter is also necessary to elicit F1 LI-ID. It is concluded that an anatomically intact spleen, i.v. presentation of antigen, and persistence of antigen within the eye are all important to the elicitation of this phenomenon.
Molecular Basis of B Cell Activation: Mitogenicity of Native and Modified Digitalis Glycosides
Authors: Hammarström Lennart L. G. , Smith C. I. Edvard
Published: December 2022

Lanatoside C, a digitalis glycoside, has previously been shown to be a potent polyclonal B cell activator for mouse lymphocytes. Since the chemical structure of lanatoside C is well defined we investigated the effect of different chemical modifications on the mitogenic properties of the glycoside in order to determine the molecular basis of B cell activation. The presence of an OH-group at C12 in the steroid nucleus proved to be essential for activation to occur in serum-free medium. Likewise, an intact carbohydrate side chain at C3 appeared to be necessary for retaining mitogenic properties. In serum-supplemented cultures, however, a much more complex response pattern was observed where the prerequisites for activation were not as distinct as in serum-free medium.
Triggering of Histamine Release from Rat Mast Cells by Divalent Antibodies against IgE-Receptors
Authors: Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

Antibodies against receptor molecules for IgE on rat basophilic leukemic (RBL) cells were prepared by immunization of a rabbit with immune precipitates composed of IgE-receptor complexes and anti-IgE. Antibodies against cell surface components were specifically purified by using RBL cells and rendered specific for mast cells by appropriate absorption. The major antibodies in the final preparation (anti-RBL) were directed against receptor molecules. It was found that the F(ab′)2 fragments of anti-RBL induced histamine release from rat mast cells and caused immediate skin reactions in normal rats. These reactions by anti-RBL or its F(ab′)2 fragments were inhibited if the receptors on mast cells had been saturated with IgE. The Fab′ fragments of anti-RBL could bind with receptors on RBL cells and blocked passive sensitization of mast cells with IgE antibodies, but failed to induce skin reactions and histamine release from normal mast cells. Sensitization of normal rat skin with the Fab′ fragments followed by an i.v. injection of anti-rabbit IgG induced skin reactions. The results indicated that bridging of receptor molecules by divalent anti-receptor antibody triggered mast cells for histamine release.
Structural and Genetic Studies on Chicken 7S Immunoglobulin Allotypes
Authors: Foppoli James M. , Wakeland Edward K. , Benedict Albert A.
Published: December 2022

Low concentrations of allotypic specificity CS-1.1 were detected in the sera of two inbred chicken lines [University of California, Davis (UCD) 7 and Regional Poultry Research Laboratory 15I4] previously reported to lack this specificty. The CS-1.1 alloantigen in 15I4 chickens has the same specificity as the major allotype in a line of chickens (UCD 2) in which it was initially defined. In 15I4 chickens, CS-1.1 allotype is present on a population of molecules distinct from those which carry the major allotype; thus a second 7S Ig H chain locus, CS-2, is proposed.

The concentration of CS-1.1-bearing molecules determined by two different methods was 7 µg/ml and 230 µg/ml in 15I4, whereas UCD 2 chickens had 4 mg/ml of CS-1.1 molecules. The levels of CS-1.1 inhibitory activity in 15I4 birds remained relatively constant over a 30-day period.

The presence of two 7S Ig populations in 15I4 chickens may be interpreted as evidence either for 7S Ig subclasses with shared allotypes or for a pseudoallelic organization of genes controlling expression of 7S Ig H chains. The results were consistent with the presence of redundant C region genes, differing in allotypes, whose expression is under the control of an as yet undefined regulatory mechanism.
Suppressor T Cells for IgE and IgG in Peyer's Patches of Mice Made Tolerant by the Oral Administration of Ovalbumin
Authors: Ngan Johnson , Kind Leon S.
Published: December 2022

A single oral dose of ovalbumin (Ov) resulted in inhibition of IgE formation in mice subsequently immunized i.p. with Al(OH)3-Ov. Repeated feeding of Ov (on alternate days for 2 weeks) induced the formation of detectable suppressor cells in Peyer's patches and spleen. Suppression was demonstrated by the ability of adoptively transferred Peyer's patch or splenic lymphocytes from Ov-fed tolerant mice to inhibit IgE formation in Ov-immunized syngeneic recipients. Suppressor cells could be induced by feeding mice as little as 100 µg of Ov on alternate days for 2 weeks. Suppression was specific and Peyer's patch lymphocytes were shown to be more effective suppressors than splenic lymphocytes.
Potentiation of Mast Cell Mediator Release by Adenosine
Authors: Marquardt Diana L. , Parker Charles W. , Sullivan Timothy J.
Published: December 2022

Since extracellular adenosine can alter adenylate cyclase activity and cell function in a variety of tissues, the effect of adenosine on mediator release was studied with isolated rat mast cells. Adenosine did not alter the rate of spontaneous release, but histamine release induced by anti-IgE, concanavalin A (Con A), 48/80, or the calcium ionophore A23187 was markedly enhanced by 1 nM to 0.1 mM adenosine. Mast cells preincubated in the presence of 10 µM adenosine and then challenged with 0.1 µg/ml of A23187 released 71.5% (± 7.4 SEM) of the cellular histamine compared to a 38.2% (± 3.0 SEM) release from control cells. Adenosine caused a 3.0-fold (± 0.2 SEM) increase in the rate of release occurring 30 to 60 sec after A23187 addition, but did not change the duration of mediator release. Adenosine did not appear to decrease the minimum concentration of ionophore required to stimulate release. The poorly metabolized adenosine analog 2-chloroadenosine (100 µM) potentiated release approximately 2-fold whereas dipyridamole, an inhibitor of mast cell uptake of adenosine, did not inhibit adenosine potentiation suggesting that adenosine actions on mast cells may be mediated by direct activation of cell-surface receptors. Adenosine modestly potentiated the fall in cyclic AMP levels occurring 30 to 60 sec after ionophore challenge, raising the possibility that potentiation may be mediated by alterations in cyclic AMP metabolism. Mast cells suspended in media containing 1 mM EGTA and no added calcium demonstrated adenosine potentiation of A23187-induced release indicating that the adenosine effect was not mediated by increased translocation of calcium into the cells. Adenosine potentiation was completely blocked by 100 µM theophylline, a known inhibitor of adenosine binding, with 50% inhibition of release occurring at 3 µM theophylline. These studies indicate that adenosine may be an important modulator of mast cell function in complex tissues and that part of the pharmacologic action of theophylline in human bronchial asthma may be through inhibition of adenosine binding, rather than direct effects on cyclic AMP metabolism.
Membrane Receptors of Mouse Leukocytes
Authors: Rabellino Enrique M. , Ross Gordon D. , Polley Margaret J.
Published: December 2022

Mouse leukocytes were studied for membrane receptors for the third C component by rosette formation with C coated erythrocytes (EAC). Methods were devised for the preparation of EAC complexes containing either mouse C3b or mouse C3d. EAC 1-3dmo were prepared from EA treated with whole mouse serum while EAC 1-3bmo were produced from EAC 142hu treated with whole mouse serum containing sodium suramin. The specificity of the EAC complexes for mouse leukocytes was confirmed by inhibition experiments using fluid phase human C3d. Low concentrations of fluid phase human C3d inhibited EAC1-3dmo rosettes but failed to inhibit EAC 1-3bmo rosettes. Eight-fold higher concentrations of fluid phase C3d caused partial inhibition of EAC1-3bmo rosette formation with lymphocytes, but not with other types of murine leukocytes. Thus mouse leukocytes apparently contain the same two types of C receptors as do human and guinea pig leukocytes. Mouse CR1 is specific for a non-C3d region of C3b, (possibly analogous to human C3c) whereas mouse CR2 is specific for both C3d and the C3d region of C3b.
Selective IgA Deficiency Associated with Total Villous Atrophy of the Small Intestine and an Organ-Specific Anti-Epithelial Cell Antibody
Authors: McCarthy D. M. , Katz S.I. , Gazze L. , Waldmann T. A. , Nelson D. L. , Strober W.
Published: December 2022

A patient with total villous atrophy of the small intestine, associated with selective IgA deficiency, presented after an acute febrile illness. Gluten-free diet and other standard treatments were without benefit. Studies revealed that T cell function, serum antibody production, delayed hypersensitivity, mixed leucocyte reaction, antibody-dependent and cell-mediated cellular cytotoxicity, and general immune responsiveness seemed intact. However, IgA could not be detected in serum, tissues, or external secretions and, cultured with poke weed mitogen, the patient's B cells failed to produce IgA. In addition, the patient's cells suppressed IgA production by control cells co-cultured in the presence of mitogen. Indirect immunofluorescent studies revealed in his serum an IgG-class organ-specific antibody which bound to villus cells in control intestine, obtained from normal and diseased subjects, but not to cells in the patient's diseased mucosa. After low dose cyclophosphamide therapy, IgA remained absent but suppressor activity disappeared, intestinal villi regenerated fully, cellular cytotoxicity was reduced, and the antibody titer in serum slowly declined. The antibody bound strikingly to mature cells in the patient's regenerated villi, on indirect but not direct immunofluorescence. These unique findings suggest an autoimmune aetiology. No such antibodies were found in other IgA-deficient subjects.
Structural Studies of Rabbit IgA Allotypes: Partial Amino Acid Sequences of α-Chain Allotypes Controlled by Alleic Genes at the Cαg Locus
Authors: Malek Thomas R. , Peterson Bruce E. , Hanly W. Carey , Knight Katherine L. , Friedenson Bernard
Published: December 2022

Four α-chain allotypic specificities of the IgA-g subclass of rabbit IgA are controlled by allelic genes at the Cαg locus. The structural basis for three of these allotypic specificities, g74, g75, and g76, was investigated by amino terminal sequence analysis. α-Chain fragments were isolated from Fc2α fragments which were obtained by tryptic digestion of g74, g75, and g76 secretory IgA. Amino acid sequence data for the chains were obtained for 92 positions; comparison of these sequences with the sequence of human α1-chain revealed approximately 70% homology with the Cα2 domain. The results indicated that g74 α-chains were cleaved 60 residues N-terminal to the cleavage site in g75 and g76 α-chains.

By comparison of sequences of 24 residues of g75 and g76 α-chains, one allotype-related difference was observed (a cys/ser interchange at position 311). Based on homologies with the human α1-chain, a model for the arrangement of disulfide bonds in rabbit α-chains is proposed; several differences between rabbit and human are indicated, as are differences among the rabbit allotypes. The most striking difference is the apparent presence of the “extra” intradomain disulfide bond in the Cα2 domain of g74 molecules and its absence in g75 and g76 molecules.
Cellular Immunity in Infectious Mononucleosis
Authors: Nikoskelainen Jukka , Ablashi Dharam V. , Isenberg Richard A. , Neel Elizabeth U. , Miller Rupert G. , Stevens David A.
Published: December 2022

Infectious mononucleosis (IM) patients, Epstein-Barr virus (EBV)-seropositive and seronegative healthy donors, and patients with other viral infections were tested for lymphocyte blastogenesis (LB) with phytohemagglutinin and six EBV (virus concentrate, culture supernatant, and soluble [S] antigen) or control antigens. Fluorescent antibodies to EBV viral capsid antigen of IgG, IgM, IgA specificities, to nuclear antigen (EBNA), and heterophile antibodies were also assayed. These were correlated with clinical parameters (fever, pharyngitis, adenopathy, hepatitis, splenomegaly, atypical lymphocytes, and total mononuclear cell counts).

EBV viral and S antigen-induced LB was significantly greater in seropositive donors. IM patients had antigen specific LB below that of seropositive donors initially and low responses for the acute phase of illness when clinical symptoms were present and antibody titers were maximal. Specific LB rose to a peak at 3.5 to 9 weeks when the patients had recovered, most laboratory findings had returned to normal, and antibodies had declined. At peak, specific LB in IM patients exceeded that of seropositive donors, but later declined.

These results demonstrate specific cell-mediated immunity (CMI) to EBV, and indicate that this develops slowly in IM and contrasts with the evolution of the clinical events and humoral immunity. This correlation supports the hypothesis that CMI is the mechanism of terminating lymphoproliferation in IM.
Radiochemical Characterization of Hemoglobins Lepore and Kenya: Unique Antigenic Determinants Located on Hybrid Hemoglobins
Authors: Garver Fred A. , Altay Gultekin , Baker Marilyn M. , Gravely Marsha , Huisman Titus H. J.
Published: December 2022

Antisera were produced in rabbits to the three known types of Lepore hemoglobins, which contain hybrid δ-β non-α-chains, and to hemoglobin Kenya, which has a hybrid γ-β non-α-chain. By using a sensitive radioimmunoassay technique, the absorbed antisera were shown to contain an antibody population that was specific for the hybrid hemoglobin and did not cross-react with normal hemoglobins. However, with the absorbed Lepore-specific antisera, the three known types of Lepore hemoglobins were antigenically indistinguishable from each other, suggesting that antibodies are not produced to the primary structural differences which define the three non-α-chains of the Lepore hemoglobins. These studies demonstrate that the non-α-subunits of hemoglobins Lepore and Kenya possess unique antigenic determinant sites, evidently resulting from an altered polypeptide conformation.
Impairment of Host vs Graft Reaction in Pregnant Mice
Authors: Clark David A. , McDermott Mark R.
Published: December 2022

The fetus resulting from an allogeneic (interstrain) mating represents a type of graft that is not rejected by the mother. Nevertheless, the maternal immune system seems to recognize and to react to the presence of the fetus in a number of ways. One such manifestation is significant enlargement of the lymph nodes that drain the uterus (DLN) of pregnant rodents. We have tested the DLN lymphocytes of mice for reactivity to paternal H-2 alloantigens after interstrain mating. The DLN lymphoid cells obtained from pregnant mice killed fewer newborn F1 recipients in a graft-vs-host mortality assay, and generated less cytotoxic T cell activity against paternal H-2 antigens both in vivo and in vitro. In vitro mixing experiments demonstrated the presence of a cell-associated suppressor activity in the DLN of pregnant mice. This suppressor proved resistant to treatment with mitomycin C, and appeared in the DLN early in pregnancy.
Release of Arylsulfatase A but Not B from Rat Mast Cells by Noncytolytic Secretory Stimuli
Authors: Lynch Susan M. , Austen K. Frank , Wasserman Stephen I.
Published: December 2022

The net percentage of release of arylsulfatase activity from purified rat mast cells induced by rabbit anti-rat F(ab′)2 was consistently only about ⅓ that of histamine. Isoelectric focusing of the released and residual arylsulfatase activities demonstrated specific release of the A type without B and a net percentage of immunologic release of arylsulfatase A equivalent to that of histamine. When the net percentage of histamine and arylsulfatase A release were nearly maximal (88 and 76%) in response to the calcium ionophore A23187, specific release of arylsulfatase B did not occur. Thus, arylsulfatase A and not B was associated with the secretory granule released from the rat mast cell by reversed anaphylaxis or the calcium ionophore. In contrast, subcellular fractionation of water-lysed mast cells yielded arylsulfatase B with the heparin- and chymase-containing granule fraction and arylsulfatase A in the aqueous fraction comprised of cell sap and granule water eluate. It may be that arylsulfatase B resides in a minor second granule, whereas arylsulfatase A is loosely associated with the predominant secretory granule of the rat mast cell.
Production of Antisera Specific for Idiotype(s) of Murine Anti-(T,G)-A--L Antibodies
Authors: Pincus Seth H. , Sachs David H. , Dickler Howard B.
Published: December 2022

Antibodies to the synthetic polypeptide (T,G)-A--L were raised in C57BL/10 and C3H.SW mice. For each strain, the anti-(T,G)-A--L antibodies from 10 animals were pooled, affinity purified on a (T,G)-A--L-Sepharose column, and used to immunize Lewis rats. The resulting rat antisera were adsorbed with insolubilized normal mouse globulin in order to remove anti-isotypic and antiallotypic antibodies. The residual antibodies specifically inhibited the binding of (T,G)-A--L by anti-(T,G)-A--L as measured by a radioimmunoassay. The specificity of this inhibition was demonstrated as follows: 1) failure of the anti-(T,G)-A--L anti-idiotype to inhibit the binding of nuclease to anti-nuclease antibody of the same allotype; 2) failure of Lewis anti-[B10 anti-(T,G)-A--L] to inhibit C3H.SW anti-(T,G)-A--L and vice versa; 3) ability to absorb anti-C3H.SW anti-idiotypic activity on insolubilized C3H.SW anti-(T,G)-A--L but not on B10 anti-(T,G)-A--L. The same or cross-reactive idiotype(s) was present in the majority of individuals of each of these strains.
Immunoregulatory Aberrations in Systemic Lupus Erythematosus
Authors: Fauci Anthony S. , Steinberg Alfred D. , Haynes Barton F. , Whalen Gail
Published: December 2022

This study was undertaken to delineate the nature of the immunoregulatory defect in patients with systemic lupus erythematosus (SLE). Thirty-eight SLE patients were studied and it was found that in addition to an absolute T lymphocytopenia, these patients had a selective proportional and quantitative deficiency (P < 0.01) of a subpopulation of T cells possessing a receptor for IgG (TG). This T cell subpopulation has been previously demonstrated to be the suppressor cell population in pokeweed mitogen (PWM)-driven intracytoplasmic Ig production. T cells with a receptor for IgM (TM) which have been shown to be the helper T cells for Ig production were not significantly different from normals in proportion or absolute numbers.

By employing a PWM-induced plaque-forming cell (PFC) assay against sheep red blood cells (SRBC), it was shown that SLE patients had a markedly reduced response of 18 (±4.2) PFC/106 lymphocytes compared to normals with 153 (±18.4) PFC/106 lymphocytes (P < 0.001). This suppressed PFC response was felt to result from in vivo polyclonal activation associated with the autoimmune state.

SLE patients had a marked defect in the ability of their lymphocytes after Con A activation to generate suppressor cells of the PWM-induced PFC response, whereas their ability to be suppressed by suppressor stimuli appeared intact. Despite considerable variability from patient to patient, lymphocytes from SLE patients in general had adequate T helper cell function and could be helped by normal allogeneic T helper cells. Thus, this study demonstrates that SLE patients have a quantitative defect in a subpopulation of T cells with suppressor capability. In addition, they have a defect in the ability to generate suppressor cell function upon appropriate activation. These findings may lend further insight into the understanding of the immunoregulatory defect in SLE.
Prolongation of Organ Allograft Survival by Syngeneic Lymphoic Cells
Authors: Tilney N. L. , Graves M. J. , Strom T. B.
Published: December 2022

The role of suppressor cells was tested in vivo by passive transfer of lymphoid cells from enhanced rats bearing well functioning cardiac allografts into unmodified syngeneic animals receiving test heart grafts 1 day later. LEW rats were enhanced with 107 BN splenocytes and 1 ml LEW anti-BN antiserum 11 and 10 days, respectively, before receiving a (LEW × BN) F1 heart graft. All such grafts function over 25 days; one-third indefinitely. Grafts in unmodified hosts survive 7 ± 0.3 days. Thymocytes (108) harvested from enhanced recipients 6 days after grafting and transferred to unmodified LEW animals prolonged test graft survival to MST of 17 ± 3 days (p < 0.001). Splenocytes (nylon wool-adherent and nonadherent fractions) from such animals prolonged test graft survival to 12 ± 4 and 14 ± 4 days, respectively. Transfer of thymocytes from immunologically virgin LEW rats or LEW rats bearing acutely rejecting (LEW × BN) F1 grafts did not prolong test graft survival. Specificity was assessed by transferring cells from enhanced LEW recipients of (LEW × BN) F1 heart grafts into LEW recipients of WF grafts. WF graft survival was not prolonged. Since increased graft survival in unmodified hosts is a stringent requirement for the effect of suppressor cells in vivo, we conclude that such cells participate in the maintenance of immunologic enhancement.
Target Antigens of Cell-Mediated Lympholysis Discrimination of HLA Subtypes by Cytotoxic Lymphocytes
Authors: Robinson Mary Ann , Noreen Harriet J. , Amos D. Bernard , Yunis Edmond J.
Published: December 2022

The evolution of HLA specificities has been toward ever-increasing refinement; one example is a subdivision of HLA-B5, the supertype specificity originally defined as 4a. HLA-B5 can now be further subdivided into Bw51 amd Bw52 by serologic means. Whereas the specificity Bw51 can be detected by specific sera, the identification of Bw52 must frequently be deduced from knowledge of B5 and Bw51, although serology has progressed rapidly. There has been no comparable development in identifying fine specificities by cellular cytotoxicity in populations. We have now found that cytotoxic effectors of exquisite specificity can be generated against Bw52 by sensitization of cells from a Bw51 donor and vice versa; Bw51 and Bw52 can in this way be recognized with equal ease. This may set a precedent for recognizing fine specificities of other HLA antigens that cannot yet be identified serologically or can be identified only imprecisely. These fine distinctions may have great relevance in allotransplantation and in understanding disease susceptibility.
The Induction of Hapten-Specific Immunologic Tolerance and Immunity in B Lymphocytes
Authors: Fidler John M.
Published: December 2022

The serum from mice that had been rendered specifically tolerant (TolS) to the trinitrophenyl (TNP) hapten by the injection of trinitrobenzenesulfonic acid (TNBS) was tested in vitro for its ability to induce unresponsiveness in untreated spleen cells. Conjugated with TNP at a concentration of ≈4 × 10-4 M, TolS induced complete TNP-specific tolerance in cultures of spleen cells challenged with TNP-lipopolysaccharide (TNP-LPS) and 4-hydroxy-3-iodo-5-nitrophenylacetic acid conjugated polymerized flagellin (NIP-POL). By using normal mouse serum (NMS) as a control, TNP-specific tolerance in the in vitro antibody response was induced by cultivation of spleen cells with amounts of TolS ranging from 10-4 to 10-2 ml (equivalent to 4 × 10-8 to 4 × 10-6 M TNP) in 1 ml cultures of 10 × 106 viable spleen cells. Thymic-derived (T) cells were not required for the induction of the unresponsiveness in TNP-specific bone marrow-derived (bursal equivalent, B) lymphocytes. TolS induced tolerance in cultures challenged with either T cell independent (TI) or T cell dependent (TD) forms of the TNP hapten. TolS was nonimmunogenic under the in vitro culture conditions used, and did not affect control responses.

Although it was most effective when harvested 4 or fewer days after TNBS injection of donor mice, TolS exerted a tolerogenic effect if obtained as much as 2 weeks after such treatment. TolS was active across both histocompatibility and immunoglobulin allotype barriers, and also exerted its tolerogenic effect in the presence of concentrations of antigen optimal for the induction of immunity. Complete tolerance was induced by pretreatment of spleen cells with TolS and extensive washing before the addition of immunogen. Following dose response kinetics, the time required and the level of unresponsiveness induced during preincubation with TolS were directly related to the dose of tolerogen utilized. The inability of TNP-lysine to act as a tolerogen and the nondialysable nature of TolS suggested that the active component of this serum is a macromolecule and not merely TNBS. Fractionation of TolS by (NH4)2SO4 resulted in virtually complete recovery of the tolerogenic activity in the globulin (nonalbumin) fraction. Although haptenated serum γ-globulins would be present in this fraction, it is also possible that other constituents with potential tolerogenic capacity are present (i.e., various serum proteins or cell surface membrane fragments that are trinitrophenylated). The data are consistent with the suggestion that TNBS-induced tolerance in vivo is mediated by a component of TolS that is active as a tolerogen in vitro.
Polymorphisms of Murine Ia Loci in Wild Mice: Serological Analysis of the I Region Antigens in a Collection of Wild Derived H-2 Haplotypes
Authors: Wakeland Edward K. , Duncan William R. , Klein Jan
Published: December 2022

The polymorphism of antigens produced by the I-A and the I-EC subregions was assessed in a collection of B10 congenic lines containing wild derived H-2 haplotypes (B10.W lines). Utilizing alloantisera prepared and analyzed with inbred lines and H-2 recombinants, a total of 29 B10.W lines were tested for 19 inbred Ia antigens of the I-A or I-EC subregions. This survey revealed: 1) that both public and private inbred Ia antigens of the I-A and the I-EC subregions are present among wild mice; 2) that the I-A subregion is more complex serologically than the I-EC region; 3) and that some wild mice do not have any of the inbred public or private antigens of either the I-A or the I-EC subregions. Serological and immunochemical characterization of alloantisera prepared in inbred lines against some of the B10.W lines has demonstrated the presence of public and private Ia antigens among wild mice that are absent from the inbred lines.
Quantitative Differences in the Expression of Specificity Ia.7
Authors: Murphy Donal B. , Jones Patricia P. , Loken Michael R. , McDevitt Hugh O.
Published: December 2022

A locus mapping between the I-J and S regions of the H-2 gene complex controls the Ia.7 determinant. Studies with recombinant strain A.TFR5 reveal that although B lymphocytes from this strain express specificity Ia.7, quantitatively less antigen is found when compared with other Ia.7-positive strains. For example, quantitative absorptions with counted numbers of lymphocytes show that roughly the same number of cells obtained from Ia.7-positive strains completely absorb antibody cytotoxic for A.TFR5 target cells from a B10.A(18R) anti-B10.A(5R) serum. In contrast, approximately five times as many A.TFR5 lymphocytes are required for complete absorption. In addition, indirect immunofluorescence studies with the same Ia antiserum show that A.TL splenic B lymphocytes stain much more brightly than A.TFR5 splenic B lymphocytes.

There are two possible explanations for these observations. First, a mutation may have occurred in the A.TFR5 line that affects the quantitative expression of the Ia.7 determinant.
Serologic Studies of I-E/I-C Gene Products in Haplotypes H-2b, H-2s, H-2q, and H-2f
Authors: David Chella S. , Neely Brian C.
Published: December 2022

The I-E and I-C subregion products have been identified specifically in haplotypes H-2k and H-2d. Haplotypes H-2p and H-2r are assumed to express products of I-E/I-C subregion on the basis of cross-reactions with anti-I-Ek sera. Anti-I-Ek or I-Ed sera do not cross-react with haplotypes H-2s, H-2b, H-2q, and H-2f. Specific immunizations to produce anti-I-Eb and anti-I-Es alloantisera have been unsuccessful. We turned to xenoimmunization to serologically identify the gene products of this chromosomal segment in the above haplotypes. NP-40 detergent solubilized antigenic material mixed in Freund's complete adjuvant were immunized into rabbits. Two weeks later a booster was given and the rabbits bled one week later for antisera analyses. The rabbits were then boosted once a month and bled periodically. The rabbit antiserum reacted with all mouse strains. Absorptions with appropriate recombinants showed the presence of specific antibodies against the I region products.
Chemical Characterization of Murine Ia Antigens
Authors: McMillan M. , Cecka J. M. , Frelinger J. G. , Murphy D. , Frelinger J. A. , McDevitt H. O. , Hood L.
Published: December 2022

Several products of the I region of the mouse H-2 complex have been characterized by micropeptide analysis, microsequence analysis, and two-dimensional gel (IEF/SDS-PAGE) analysis. Ia antigens have been isolated using the biosynthetic radiolabeling technique together with highly specific alloantisera and indirect immunoprecipitation. Chromatography of tryptic peptides prepared from the α and β polypeptides of Ia antigens encoded by the I-A and I-EC subregions will be compared from both spleen and epidermal cell preparations. These data, in conjunction with partial N-terminal amino acid sequence data on Ia antigens from the I-Ab, I-Ak, I-Ad, I-As, I-ECk and I-ECd subregions and haplotypes will be discussed in terms of homology relationships and haplotype-associated differences in the structures of these Ia polypeptides. Two-dimensional gel patterns of isolated α and β polypeptides will also be presented. The implications of these data will be discussed as they relate to the genetic organization of the I-region.
Isoelectric Focusing of Guinea Pig Ia Antigens
Authors: Schwartz Benjamin D. , Gordon Diane , Thomas David W.
Published: December 2022

To ascertain whether guinea pig Ia antigen-bearing molecules might act as receptors for exogenous antigens, Ia molecules from strain 2 and strain 13 guinea pigs were tested for the presence of variable regions by isoelectric focusing (IEF). Ia antigens from lymph node cells, T lymphocytes, and macrophages were radiolabeled with 3H-leucine or 35S-methionine, solubilized by NP-40, and isolated by immunoprecipitation. The IEF characteristics of each component Ia chain were studied by employing a “reverse” O'Farrell technique whereby the two Ia chains were first separated by SDS-polyacrylamide gel electrophoresis, and then each chain was subjected to IEF. Comparison of Ia molecules derived from each of the cell populations, but bearing identical Ia determinants, revealed virtually identical IEF patterns, suggesting that the same class of Ia molecules can be derived from each cell population studied. Within each inbred strain, Ia molecules bearing different determinants had similar but distinctive IEF patterns.
Complex Genetic Control of Immune Response to Lysozymes in Mice
Authors: Kipp Dale E. , Miller Alexander , Sercarz Eli E. , Taylor Benjamin A.
Published: December 2022

H-2b mice fail to respond to some bird lysozymes (prototype: hen egg-white lysozyme (HEL)) while showing normal responsiveness to other closely related bird lysozymes (prototype: ringed-neck pheasant lysozyme (REL)). H-2b mice also fail to respond to human lysozyme (HUL). Previous studies have shown that differential HEL:REL responsiveness is associated with two genetic loci within H-2, one mapping to the left of I-B and one to the right. Present studies, in which equal positive response is found to HEL and REL in H-2ba and H-2bf mice, indicate that a mutation at H-2K is associated with the loss of ability to discriminate between HEL and REL. The presence of at least one controlling locus outside H-2 is suggested by studies with recombinant inbred strains (RI) of C57BL/6 and C57L (a strain which responds to HEL although H-2b). Two such RI with H-2 derived from the C57BL/6 parent respond to HEL while one RI with H-2 derived from C57L does not respond.