bluuebunny/crossref_metadata_2025 huggingface dataset:
Alteration in Complement Component Utilization Following the Interaction of C1̄hu with a Kallikrein Fragment
Authors: Gigli I. , Kaplan A. , Austen K. F.
Published: December 2022

A mixture of human IgG, kallikrein and plasma thromboplastin antecedent (PTA) prepared by passage of serum over DEAE-cellulose equilibrated with 0.01 m phosphate buffer, pH 8, was found to enhance the hemolytic activity of activated human first component of complement (C1̄hu). Fractionation of this mixture on Sephadex G-150 revealed peaks of C1̄hu-enhancing activity at 45% bed volume, the position of kallikrein, and at 80% bed volume. Repeat filtration of the kallikrein fraction on Sephadex G-150 yielded the low molecular weight C1̄hu-enhancing factor, suggesting that it was derived from kallikrein. This non-dialyzable fragment lacked kallikrein activity and unlike C1̄s did not split N-α-acetylglycyl-l-lysine methylester (AGLMe), N-acetyl-l-arginine methylester (AAMe), p-tosyl-l-arginine methylester (TAMe), N-acetyl-l-tyrosine methylester (ATMe), the fourth component (C4), or the second component (C2).

Kinetics of C2 activation by C1̄ acting upon EAC42 prepared with limited concentrations of unactivated C2 indicated that C1̄ containing 20 mol/cell produced 0.57 SAC with a tmax of 3.5 min; C1̄ acted upon by the fragment and then diluted sevenfold as compared to the untreated C1̄ produced 1.6 SAC at a tmax of 1.75 min.
C4 Inactivating Factor in C4 Deficient Human Serum
Authors: Torisu M. , Nagaki K. , Inai S. , Sonozaki H.
Published: December 2022

Our inactivating factor for the fourth component of human complement (C4 INF) was partially purified from pseudoglobulin fraction of C4 deficient human serum. Initially C4 INF was thought to specifically inactivate C4; however, after thorough comparison with highly purified active human esterase (C1s), it became clear that both substances share the following activities: 1) Both substances showed SAC4,2 generating activities on EAC4gp cells as well as on EAC4hu cells. 2) Both substances inactivated C2 as well as C4. 3) Both substances hydrolyzed TAMe. 4) Both substances were sensitive to DFP as well as PMSF. 5) SAC4,2 generating activities of both substances were neutralized by anti-C1s. 6) Both substances were not fixed on either EA or EAC4 cells. 7) Both substances showed antigenic identity against anti-C1s by Ouchterlony analysis. 8) Both substances showed the same electrophoretic mobility on acrylamide gel. 9) Both substances altered the electrophoretic mobility of native C4hu in a similar manner.
Isolation and Characterization of a Factor from Human and Guinea Pig Serum That Accelerates the Decay of SAC142
Authors: Opferkuch W. , Loos M. , Borsos T.
Published: December 2022

A factor has been isolated from the pseudoglobulin fraction of human and guinea pig serum which accelerates the decay of SAC142. The decay-accelerating factor (DAF) of guinea pig serum is a heat stable (1 hr, 56°C) α globulin, has a molecular weight of about 300,000 (estimated by gel filtration) and a sedimentation rate of 8.2S; the DAF of human serum has an m.w. of about 170,000. DAF can be adsorbed to E at a rate that is independent of temperature. EA, EAC1, EAC4, EAC14 and EAC142 at the same cell concentration take up the same amount of DAF. The activity of a given amount of DAF is independent of the number of SAC142/EAC142 and is dependent on the number of cells present in the reaction mixture. While DAFhu accelerates the decay of SAC142 made with C2hu, oxyC2hu and C2gp, DAFgp is capable of accelerating the decay of SAC142gp, but not those made with C2 of human origin.
Isolation of the Precursor and the Active Form of the C3 Activator from Human Serum
Authors: Götze O. , Müller-Eberhard H. J.
Published: December 2022

It had previously been shown that a protein isolated from the venom of Naja naja, the cobra factor, is capable of forming a complex with a β globulin present in human serum. This complex is able to cleave C3 in free solution and apparently activates the terminal complement components beginning with C3.

The C3 proactivator (C3PA) which participates in these reactions by forming the enzymatically active complex with cobra factor was isolated from human serum. It was found to be homogeneous on disc- and immunoelectrophoresis.

Cleavage of the C3PA into at least two fragments was observed by treatment of serum with a variety of naturally occurring lipopolysaccharides and polysaccharides such as endotoxin, zymosan, inulin or agar. This reaction was accompanied in all instances by the conversion of C3 to its electrophoretically faster form. The larger of the two C3PA fragments (m.w. approximately 60,000) has γ globulin mobility and is antigenically deficient.
Immunochemical and Biologic Characterization of a New C3 Conversion Product, C3x, Generated in Human Serum in the Absence of C3 Convertase
Authors: Spitzer R. E. , Stitzel A. E. , Pauling V. L. , Davis N. C. , West C. D.
Published: December 2022

C3x, the product of a non-convertase dependent C3 conversion mechanism in normal human serum, appears to be a conformational alteration of the native molecule. It has a molecular weight approximately the same as C3 and its generation is not accompanied by any detectable cleavage of the molecule. C3x bears all three antigens of C3 (A, B, D) but is quantitatively deficient with respect to the native molecule in the amount of antibody consumption to the B antigen. In addition, C3x has a slightly faster mobility in acrylamide gel and agar than does native C3. C3x is extremely stable, resisting cleavage by trypsin up to 1 hr at 37°C, 0.3 M hydroxyl-amine, C, or pH 3.0. Biologically, C3x is unique in that it cannot participate in immune lysis or erythrocyte immune adherence but does interact with C5 and augments erythrophagocytosis. C3x is also markedly chemotactic for human PMN's and causes increased vascular permeability in the guinea pig which is not histamine dependent.
Complement Efficiency in Immune Hemolysis
Authors: Linscott W. D.
Published: December 2022

C1 is fixed very inefficiently by IgG anti-Forssman antibody under physiologic conditions of temperature and ionic strength (µ), and these are the same conditions under which most hemolytic and C fixation reactions are carried out. If washed, IgG-sensitized erythrocytes are incubated with absorbed guinea pig C for 1 hr at 0°C, µ = 0.037, and then diluted with an equal volume of cold buffer at µ = 0.15 and immediately placed at 37°C for 1 hr, a more than 10-fold increase in sensitivity of the assay is obtained (less IgG and less C are needed for hemolysis).

The sensitivity of hemolytic assays for IgM can also be improved, though not so dramatically, by choosing conditions more favorable for C1 fixation. At low antibody concentrations, a severalfold increase in lysis is observed if washed, IgM-sensitized erythrocytes are incubated with absorbed guinea pig C for 1 hr at 37°C, µ = 0.094, instead of µ = 0.15.
Ultrastructural Studies of Clq and of the Interaction Between Membranes and Complement Proteins
Authors: Polley M. J.
Published: December 2022

Ultrastructural studies of human C1q have demonstrated the molecule to exist in two distinct forms. Svehag and Bloth have described the molecule to be composed of six subunits arranged in a disc-like form. Our studies suggest that the disc-like form of the native molecule constitutes a disc-like coil consisting of six subunits which is capable of unfolding to a rod-like structure with an identical number of subunits.

Red cell antibody complement complexes were prepared at various intermediate stages throughout the complement sequence; stroma was prepared from each and examined ultrastructurally. In complexes prepared up to the EAC1,4,2,3 stage in which no lesions were seen, sections of the membranes demonstrated the “tram-line” appearance described for normal red cell membranes by Dourmashkin and Rosse. At the EAC1–5 stage, when lesions first became visible, the membranes seen in section had retained their dense linear appearance; however, they had increased in width from 100 to 250 Å.
Kinetics of Bovine Complement
Authors: Fong Jack S. C. , Muschel Louis H. , Good Robert A.
Published: December 2022

With the use of conglutination assay and different anticomplementary agents, the lytic intermediate formed during interaction of sensitized sheep erythrocytes and diluted bovine serum was shown indirectly to be SHEACBo142. Lysis of this cellular intermediate by guinea pig C3-9 was completed in about 6 min of incubation at 37°C at ionic strengths between 0.075 and 0.125. In contrast to the lability of other corresponding intermediates, SHEACBo142 was extremely stable in low ionic strength buffer (GGBS++) with divalent cations: up to 18 hr at 37°C and 6 days at 2°C. Change of the ionic strength of the buffer to iso-ionicity resulted in gradual decay of the lytic intermediate, whereas presence of EDTA induced a precipitous decay of SHEACBo-142, whose reactivity could be restored with bovine euglobulin or functionally pure human C1. The apparent paradox, that is, that bovine complement was non-hemolytic to sensitized sheep erythrocytes and inhibitory to the lytic activity of guinea pig terminal complement components on SHEACBo142 but hemolytic to sensitized rabbit erythrocytes, further emphasized that in vitvo hemolytic complement assays do not necessarily correlate with in vivo functional activity of complement-dependent or -mediated processes.
Lack of Antibody Activity in Human Myeloma Proteins
Authors: Yoo T. J. , Franklin Edward C.
Published: December 2022

Two hundred and seventy-five myeloma sera were screened with a battery of 20 antigens including 13 haptens, pneumococcal polysaccharides and four bacterial antigens. In no instance was it possible to detect antibody activity by double diffusion in agar or by fluorescence quenching.
Surface Immunoglobulins on Mouse Lymphoid Cells
Authors: Bankhurst Arthur D. , Warner Noel L.
Published: December 2022

Addition of 125I-labeled anti-immunoglobulin protein to cell suspensions from mouse thymus, spleen and thoracic duct lymph (TDL) resulted in labeling of surface immunoglobulins of lymphoid-like cells. Near-optimal conditions for cell labeling were established with varying anti-immunoglobulin concentrations and times of exposure. On the average 20.1% to 36.3% of TDL and spleen cells labeled under such conditions, whereas thymus cells labeled less heavily (0.3% to 3.7%) with identical materials. Normal rabbit sera (NRS) or anti-immunoglobulin absorbed with purified immunoglobulins produced essentially no labeling. The total heavy chain labeling on TDL and spleen cells exceeded light chain labeling (64.6% vs 31.8% and 79.8% vs 33.0%, respectively). This might suggest that labeled cells had multiple surface heavy chain determinants or that the anti-heavy chain antibodies cross-reacted with an unidentified immunoglobulin-like surface antigen. There was increased labeling with TDL suspensions from neonatally thymectomized mice (65% with anti-light chain). This was consistent with the hypothesis that primarily non-thymus-derived cells were labeled by these techniques.
An Efficient Method for Isolation of HL-A Antigens from Hematopoietic Cell Lines
Authors: Miyakawa Y. , Tanigaki N. , Yagi Y. , Pressman D.
Published: December 2022

A simple isolation procedure has been established that gives a high yield of relatively pure HL-A antigens from cells of established hematopoietic cell lines. Particulate HL-A antigens were obtained from cells disrupted by shaking in buffer. These particulates were digested with papain in the presence of cysteine, and the soluble papain fragments that carried antigenic activity were purified by hypotonic dialysis, followed by ion exchange chromatography on DEAE-Sephadex, ultrafiltration, gel filtration and column electrophoresis. Several preparations of soluble HL-A antigens, each carrying one of the representative antigenic activities, HL-A1, HL-A2 or HL-A7, were obtained from three different cell lines. These preparations had a high degree of homogeneity in molecular size (48,000 Daltons) and a narrow range of electrophoretic mobility in the α2 globulin region. In these preparations, the overall recovery of HL-A antigenic activity was 30% to 37% for each isolated specificity, the recovery of OD280 units was 0.08% to 0.14%, the OD280/OD260 ratio was approximately 1.7 and the increase in specific antigenic activity was approximately 300 times that of papain digests.
HL-A Antigens from Hematopoietic Cell Lines: Molecular Size and Electrophoretic Mobility
Authors: Tanigaki N. , Miyakawa Y. , Yagi Y. , Pressman D.
Published: December 2022

HL-A antigens of hematopoietic cells in long-term culture were solubilized by papain digestion, and the molecular size and electrophoretic mobility of the fragments carrying individual HL-A specificities were estimated. HL-A1, HL-A2, HL-A7 and HL-A8 specificities were present on molecular fragments of 48,000 Daltons. An unidentified alloantigenic specificity was present on molecular fragments of 74,000 Daltons. Although all the HL-A active fragments migrated to the α2 globulin region upon electrophoresis, the mobility of the molecular fragments carrying HL-A2 specificity was slightly lower than those of fragments having the other specificities. All papain fragments of HL-A2 specificity from different cell lines were identical in molecular size and electrophoretic mobility. A similar identity also appeared to be present among the papain fragments carrying the other specificities.
Rabbit Secretory IgA
Authors: O'Daly Jose A. , Cebra John J.
Published: December 2022

Rabbit free secretory component was isolated from colostrum and some of its molecular characteristics were compared with those of secretory component dissociated from sIgA. The specific in vitro association of free secretory component with sIgA rendered deficient in bound secretory component is described.
The Immunosuppressive Effects of Rauscher Leukemia Virus Upon Spleen Cells Cultured in Cell-Impermeable Diffusion Chambers
Authors: Borella Luis
Published: December 2022

Qualitative and quantitative differences were demonstrated in the effects of Rauscher leukemia virus (RLV) upon various expressions of the immune response. BALB/c mouse spleen cells were inoculated with RLV and cultured in cell-impermeable diffusion chambers. The suppressive effects of RLV on the cellular expression of IgM and IgG memory and on the cell responses to phytohemagglutinin (PHA) and alloantigens were investigated. RLV inoculated at time zero of culture inhibited IgG but not IgM memory. Both responses were depressed when the interval between RLV inoculation and antigenic restimulation was increased. Removal of the macrophage-rich, adherent cell population depressed the IgM but not the IgG response, indicating that the early selective inhibition of IgG memory was not dependent on infection of macrophages by RLV. Incorporation of 3H-TdR was higher in cultures of spleen cells infected with RLV than in non-infected controls. This high background precluded accurate measurement of stimulation of DNA synthesis by RLV-infected BALB/c cells cultured with mitomycin-treated C57BL mouse spleen cells. However, infected cultures responded to PHA stimulation with 3H-TdR incorporation similar to that of the non-infected, PHA-stimulated cells. These data indicated that RLV-induced immunosuppression is not an all-or-none phenomenon; the degree of inhibition appeared to depend on the different immunocompetent units participating in various immune responses.
Role of the Bursa of Fabricius in IgG and IgM Production in the Chicken: Evidence for the Role of a Non-Bursal Site in the Development of Humoral Immunity
Authors: Lerner K. G. , Glick B. , McDuffie F. C.
Published: December 2022

Ablation of the bursa of Fabricius in the chicken at different times and by different techniques was carried out to assess the relative effects on production of antibodies to SRBC and of IgM and IgG. In confirmation of previous work, IgG and IgG antibody production were readily inhibited by bursectomy. In contrast, IgM production was increased by the methods used unless irradiation was added. Dipping eggs in testosterone propionate at 3 days of incubation, which completely prevented development of the bursa, resulted in IgM levels at 14 wk that were 4 × those of controls but not accompanied by any increase in the level of IgM anti-SRBC. The results indicate that: 1) the bursa has a major role in the maturation from IgM to IgG synthesis, and 2) the bursa is not essential for antibody and immunoglobulin production.
A Rapid Method for Direct HL-A Typing of Cultured Lymphoid Cells
Authors: Ferrone Soldano , Pellegrino Michele A. , Reisfeld Ralph A.
Published: December 2022

Extensive use of cultured human lymphoid cell lines in transplantation immunology has increased the need for a simple and rapid typing method to directly determine their HL-A phenotype. Such a procedure would also permit the use of phenotyped cultured cell lines as a standard panel, thus improving the screening and characterization of cytotoxic HL-A alloantisera and facilitating comparison of these sera among different laboratories. Furthermore, since cultured cells are an excellent source of soluble HL-A antigens (1, 2), a rapid and direct method to determine their mosaic of HL-A determinants would facilitate the isolation and biologic characterization of these antigens. Thus far, cultured cells cannot be used efficiently in HL-A typing laboratories since these cells are extremely sensitive to the natually occurring cytotoxins in rabbit serum (3, 4) that is routinely employed as a source of complement (C) in the C-dependent cytotoxic reaction.
Characterization of Three Homogeneous Canine Immunoglobulins
Authors: Hurvitz Arthur I. , Kehoe J. Michael , Capra J. Donald
Published: December 2022

Three homogeneous immunoglobulins were isolated from the serum or urine of three dogs with plasma cell dyscrasias. Antigenic and physicochemical characterization of the proteins showed that one was a macroglobulin, a second a γA myeloma paraprotein, and the third a Bence Jones protein. The proteins were characterized by physical, chemical and immunologic analysis and compared with human immunoglobulins, as well as with a previously studied canine macroglobulin. The studies confirmed the production of myeloma paraproteins by certain dogs with plasma cell dyscrasias and suggested their suitability for further elucidating the nature of canine immunoglobulins. In addition, such pure proteins should prove useful for comparative amino acid sequence analyses with human and murine myeloma proteins.
Activity in Vitro of Lymphocytes and Macrophages in Delayed Hypersensitivity
Authors: Salvin S. B. , Sell Stewart , Nishio Jane
Published: December 2022

The in vitro activities and interactions of macrophages and lymphocytes from tuberculin-sensitive and normal guinea pigs, in the presence or absence of antigen (PPD), were examined with the aid of phase-contrast time-lapse cinematography. Macrophages from peritoneal exudate had active amoeboid motion and numerous rapidly extending pseudopodia. Macrophages from normal or sensitized animals, in the presence of antigen and sensitized lymphocytes, and to a lesser extent sensitized thymocytes, formed considerably less active and less numerous pseudopodia, became spherical and were more sluggish in their movements. The motion of lymphocytes and thymocytes, normal or sensitized, was not affected by specific antigen. Non-sensitized macrophages aggregated and tended to become associated with lymphocytes in the presence of antigen and sensitized lymphocytes. These in vitro observations provided direct evidence of immunologically induced cellular interactions in delayed hypersensitivity systems.
Isolation and Purification of a Peptido-Rhamnomannan from the Yeast Form of Sporothrix Schenckii. Structural and Immunochemical Studies
Authors: Lloyd Kenneth O. , Bitoon Minda A.
Published: December 2022

A peptido-rhamnomannan was isolated from the cells and culture medium of the human pathogenic fungus Sporothrix schenckii, grown in the yeast phase. It was purified by precipitation of its borate complex with hexadecyltrimethylammonium bromide and then by DEAE-Sephadex chromatography. The sugars in the polysaccharide were identified as d-mannose (50%) and l-rhamnose (33%). The amino acid composition of the peptide resembled that of the peptide moiety of other fungal peptido-polysaccharides. The polymer was almost entirely (91%) precipitated by concanavalin A and the ratio of mannose to rhamnose in the precipitate was unchanged, showing that the sugars are components of the same polymer. Methylation analysis and partial acid hydrolysis showed that the basic structure of the polysaccharide is a peptido-mannan to which numerous l-rhamnose residues are attached. About 60% of the rhamnose residues could be preferentially removed by mild acid hydrolysis. The peptido-rhamnomannan precipitated with the sera of individuals (3) with sporotrichosis. Sera from a small number of patients with other fungal diseases and from normal individuals did not precipitate within the sensitivity limits of the method. The specificity of two of the sera were examined in more detail by studying their cross-reactions with other fungal mannans.
Augmented Anti-SIII Antibody Responses to an SIII-Protein Conjugate
Authors: Paul William E. , Katz David H. , Benacerraf Baruj
Published: December 2022

Rabbits that had been primed with type III pneumococci (PnIII) manifested striking anamnestic anti-type-specific polysaccharide (SIII) responses to challenge with a covalent conjugate of SIII and bovine γ globulin (BGG) if they had received an intervening immunization with BGG. If no supplemental BGG immunization was given, the SIII-BGG complex caused no secondary anti-SIII response in primed animals. Furthermore, a single intravenous dose of SIII-BGG complex elicited a primary anti-SIII antibody response in rabbits. These results were interpreted in the light of cooperative cellular interactions in immune responses. An approach of this sort may be suitable for clinical immunization against a variety of bacterial polysaccharides.
Spontaneous Reoxidation of the Subunits of Immunoglobulin M (IgM)
Authors: Mukkur T. K. S. , Inman F. P.
Published: December 2022

The consequences of subsequent reoxidation of human IgM that had been lightly reduced or extensively reduced were studied. When IgM was reduced with 0.02 M mercaptoethylamine (MEA), only the intersubunit interchain disulfide bonds were broken. The interchain bonds within each subunit, IgMs, remained intact. The percentage of μ and L chains in the alkylated IgMs, as measured by absorbancy at 280 nm, was virtually the same as that of the parent IgM (77% μ; 23% L). Reoxidation of the MEA-produced IgMs, in the absence of alkylation, led to the formation mainly of oxidized residual IgMs and apparently IgM. The percentage of μ and L chains in the reoxidized IgMs was the same as that of alkylated IgMs. In addition, the sedimentation coefficients of alkylated and of reoxidized IgMs were identical. On the other hand, reduction of IgM with 0.1 M 2-mercaptoethanol (2-ME) resulted in cleavage of all interchain bonds in the molecule. In the absence of alkylation, spontaneous reoxidation of the mixture led to the formation of several entities intermediate in size between IgMs and IgM. The percentage of L chains, again measured by absorbance at 280 nm, in the alkylated 2-ME-produced IgMs and in reoxidized IgMs was a little lower than that of parent IgM; that of one of the faster sedimenting reoxidation products was only 12%. It was concluded that the nature of the oxidation products depended upon whether only intersubunit disulfide bonds were split or whether there also was extensive cleavage of interchain disulfide bonds within the subunits.
Antigenic Competition between Mouse Thyroglobulin and Bovine Serum Albumin Demonstrated by Adoptive Transfer
Authors: Twarog Frank J. , Rose Noel R.
Published: December 2022

Antigenic interaction was found when lymphoid cells were transferred from donors immunized with both mouse thyroid extract (MTE) and bovine serum albumin (BSA) and boostered 3 days before transfer. BSA antibody was suppressed and MTE antibody was enhanced in recipients of spleen and lymph node cells from these donors.
Detection of Bound C3 by a New Immunochemical Method
Authors: Borsos Tibor , Leonard Edward J.
Published: December 2022

C3 bound to sheep erythrocytes (EC3) reacts with anti-C3 antibody. The C3-anti-C3 complex fixes complement and can be detected quantitatively by either 1) the C1 fixation and transfer test or 2) the amount of lysis which occurs when whole complement is added to the erythrocyte-C3-anti-C3 complex. The hemolytic activity of anti-C3 antibody is inhibited by fluid phase C3 or by C3 bound to immune complexes. We have devised a quantitative test for C3 antigen based on the inhibition of anti-C3 antibody by C3. The C3 to be measured (in either bound or fluid phase form) is added to a standard amount of anti-C3 antibody. Residual antibody activity is then determined by adding the absorbed antiserum to sheep EC3 indicator cells and measuring lysis of the cells which occurs after addition of whole guinea pig complement. By comparing the observed lysis with an inhibition-of-lysis curve obtained with known C3 antigen, the amount of test antigen can be estimated. The method is simple, rapid and reproducible and can detect nanogram amounts of bound C3.
Evolution of the Immune Response
Authors: Litman G. W. , Frommel D. , Finstad J. , Good R. A.
Published: December 2022

The subunit nature of bowfin (Amia calva) immunoglobulins was examined by a variety of physicochemical techniques. Dithiothreitol (DTT) reduced the 13.6S bowfin immunoglobulin, thought to be tetrameric, into monomeric subunits. The limited reduction procedure, designed to reduce only inter-subunit disulfides, cleaved light chains from the subunits and resulted in loss of the parent molecule's ability to agglutinate Brucella abortus. Relatively weak non-covalent bonding was detected between the heavy and light chains of both the high and low molecular weight bowfin immunoglobulins. The isolated bowfin heavy and light chains possessed CD spectra not unlike those of human μ heavy chains and κ light chains examined under identical solvent conditions. A mass difference, not attributable to carbohydrate, of ∼ 12,000 was detected between the heavy chains of the two size classes of bowfin immunoglobulin. When the heavy chains were compared antigenically, no differences were detected. Mass deletions of this nature lent support to the hypothesis that immunoglobulins evolved by end-to-end duplication of a primordial gene coding for approximately 110 amino acids.
Inhibition of Macrophage Migration: A Test System Using Human Monocytes
Authors: Goldberg Leonard S. , Louie James S. , Baker Mary H.
Published: December 2022

A method for the assay of macrophage migration inhibiting factor (MIF) using human lymphocytes and isolated human peripheral blood monocytes is described. This technique allows one to investigate the lymphocyte's ability to produce MIF and the monocyte's ability to respond to it.
Studies with Anti-Pneumococcal Antibodies
Authors: Haber E. , Jaton J. C. , Waterfield M. D. , Margolies M. N. , Bloch K. J. , Cheng W. C.
Published: December 2022

Antibodies from a single rabbit immunized with type VIII pneumococci were isolated after each of three courses of immunization. A common amino acid sequence was demonstrable in the N-terminal region in all three antibody light chain preparations. This was the predominant sequence detected in second course antibody (>90%); additional residues at several positions, were found in the first and third course antibodies. Peptide maps and radioautographs of chymotryptic digests of light chains, reduced and alkylated with [2-14C]-iodoacetic acid revealed two major cysteine-containing peptides in all three preparations. These peptides originated from the variable region of the light chain and were found in digests of non-antibody immunoglobulins of the same allotype. In the first and third course antibody, these two peptides were present in lower concentrations than in the second course antibody and were accompanied by additional cysteine-containing peptides.
Sequence Studies on Rabbit γG Heavy Chains
Authors: Fleischman J. B.
Published: December 2022

Amino acid sequence studies in certain regions of rabbit IgG heavy chains have been hampered by heterogeneity of the material. To overcome this, an antibody with uniform properties to group C streptococcal carbohydrate was isolated from the serum of a single rabbit. The antibody was mainly of a2 allotype; it showed restricted electrophoretic mobility and was more than 90% precipitable with group C carbohydrate. A partial sequence of 65 residues was obtained in the N-terminal part of its heavy chain, including a section hitherto unreported in any rabbit IgG. A comparison of the a2 sequence with those of the a1 and a3 allotypes revealed that about half of the first 34 residues are identical in all three allotypes.
Amyloid Proteins: The Amino-Terminal Variable Fragment of Immunoglobulins
Authors: Glenner G. G. , Harada M. , Isersky C. , Potter M. , Page D.
Published: December 2022

Purification of the major protein component of amyloid fibrils from human tissues and from tissues of the experimental murine disease has been accomplished by sequential gel filtration on Sepharose 4 B and Sephadex G-100 columns using 5 M guanidine-HCl in 1 N acetic acid with removal of over 28% of minor constituents. All amyloid proteins have a constant tryptophan content, a high dicarboxylic and short chain amino acid composition, an unreactive or Asx N-terminal group, monomeric molecular weight ranges of from 5000 to 18,300 and, in the human, heterogeneity of amyloid protein between individuals and homogeneity within the individual. Antibodies directed against the major amyloid protein of an individual cross-react with a component in the patient's serum. Immunologic evidence reveals the experimental murine protein to be derived from the N-terminal fragment of an immunoglobulin light chain of λ type and sequence analysis of a human protein categorizes it as derived from the N-terminal variable fragment of a κ I light chain.
Immune Cell Function, Viruses and Autoimmunity in New Zealand Mice
Authors: Talal Norman , Steinberg Alfred D. , Jacobs Michael E. , Gazdar Adi
Published: December 2022

The development of autoimmunity in New Zealand mice is related to genetic, immunologic and viral factors. Evidence was presented to suggest that thymus-dependent immune functions may be depressed and bone marrow-dependent functions augmented in these mice. Antibodies to RNA and DNA appear spontaneously and can also be induced by treatment with polyinosinic·polycytidylic acid (rI·rC). Antibodies binding 14C-rI·rC in human lupus sera and in New Zealand mice developing lupus have greatest specificity for reovirus double-stranded RNA. Treatment of New Zealand mice with RNA and cyclophosphamide induces immunologic tolerance and suppresses antibodies binding 14C-rI·rC. Induction of tolerance in this way prevents the accelerated formation of anti-RNA antibodies normally induced by Moloney leukemia virus. This finding suggests that virus-accelerated and natural disease occur through a similar mechanism, and supports the hypothesis that common viruses may act as an antigenic stimuli for a genetically hyper-responsive antibody-producing system.
Procholeragenoid: An Aggregated Intermediate in the Formation of Choleragenoid
Authors: Finkelstein Richard A. , Fujita Koichiro , LoSpalluto Joseph J.
Published: December 2022

A high molecular weight polymer, designated “procholeragenoid,” is an intermediate in the heat-mediated conversion of the cholera exo-enterotoxin, choleragen, to the natural toxoid, choleragenoid. The demonstration of this polymer offers an explanation for the conversion of the toxin, ca. 90,000 m.w., to the antigenically identical toxoid, ca. 60,000 m.w., without apparent loss of a fragment. Isolated procholeragenoid has little residual toxicity and is immunogenic for mice.
Suppression of Antibody Synthesis
Authors: Fauci Anthony S. , Johnson John S.
Published: December 2022

Antigenic competition between two specific haptens was demonstrated at the cellular level in local rabbit lymph nodes. Competition was shown to exist whether the hapten was linked to the same or to different carriers. Competition existing at certain critical doses of immunogen indicated that the phenomenon is not a true competition for a cell type or for some aspect of the antigen-processing system. The test antigen and the competing antigen were injected simultaneously into opposite side footpads of the rabbits and suppression was demonstrated in local lymph node cell responses. The data indicate that a circulating inhibitor was elaborated during the immune response.
Graft-vs-Host Inhibition
Authors: Bortin Mortimer M. , Rimm Alfred A. , Saltzstein Edward C.
Published: December 2022

An in vitro treatment of immunocompetent cells was tested in an effort to prevent graft-vs-host (GVH) disease and to promote survival of allogenic radiation chimeras. Spleen cells from adult strain-A mice were mixed and incubated for 1 hr at 37°C with liver cells from 5-day-old strain-A mice. There was no apparent decrease in hematopoietic stem cell function in the incubated liver and spleen cell mixtures when compared with stem cell function in separate cell suspensions maintained at 4°C. Severe GVH disease occurred when fresh strain-A spleen cells were given to lethally irradiated CBA mice. No mice survived longer than 33 days. Incubation of spleen cells alone for 1 hr at 37°C or incubation of liver-spleen cell mixtures for 1 hr at 4°C did not appreciably improve survival. Incubation of spleen cells alone for 2 hr at 37°C caused improved 30-day survival of the allogeneic radiation chimeras. Significant inhibition of GVH disease was also observed when lethally irradiated CBA mice were treated with strain-A spleen cells that had been incubated with strain-A liver cells for 1 hr at 37°C.
Immunity to Disease Caused by Hemophilus Influenzae Type b
Authors: Schneerson Rachel , Rodrigues Louis P. , Parke J. C. , Robbins John B.
Published: December 2022

Purified type b capsular polysaccharide of Hemophilus influenzae induced a serum antibody response in 20 adults with pre-existing antibodies. Using absorption experiments in mouse protection and passive hemagglutination and complement-dependent bactericidal assays, the anti-type b antibodies induced by immunization had identical characteristics to “natural” and infection-induced antibodies. Further evidence for the protective quality of anti-type b antibodies was derived from the finding of an inverse relationship between the age incidence of influenzal meningitis and the percentage of individuals of different ages with serum anti-capsular antibodies. Most infants and children with disease did not have anti-capsular antibodies during the acute phase of their disease. Patients with antibody deficiency syndromes, highly susceptible to disease with H. influenzae type b, did not have detectable serum anti-type b antibodies.
Removal of Neuraminic Acid from H-2 Alloantigens Without Effect on Antigenic Reactivity
Authors: Shimada Akira , Nathenson Stanley G.
Published: December 2022

Murine H-2 alloantigen glycoprotein fragments (10% carbohydrate comprised of sialic acid, fucose, galactose, mannose and glucosamine) were treated with Vibrio cholerae neuraminidase. While nearly 100% of the sialic acid was removed, no loss of H-2 antigenic reactivity occurred.
Functional Relationship of Factor B in the Properdin System to C3 Proactivator of Human Serum
Authors: Goodkofsky Ira , Lepow Irwin H.
Published: December 2022

Factor B in the properdin system is a heatlabile protein requiring, in addition to properdin, other serum factors, and Mg2+, for formation of a complex with zymosan which preferentially inactivates C3 and later acting components. Factor B is a β-pseudoglobulin which is distinct from C2. Evidence has been presented that factor B is functionally identical to an independently isolated serum protein, C3 proactivator (C3PA). These observations serve both to identify and characterize factor B and, in conjunction with earlier evidence, to establish the properdin system as an alternate pathway to C3 and later acting components, independent of C1, C4 and C2.
Peptides from the Site of Rabbit Antibody: Anti-3-Azopyridine Antibody
Authors: Friedenson Bernard , Takeda Yasushi , Roholt Oliver A. , Pressman David
Published: December 2022

The amino acid sequences of two peptides containing tyrosine that appears to be derived from the combining site of an anti-3-azopyridine antibody were determined. Binding studies showed that the antibody was inactivated by iodination and that the attack was in the site, since the presence of the hapten, pyridine, during iodination prevented the loss of sites. Peptides from the site were isolated after a paired labeling procedure as follows. One portion of the antibody was iodinated with 125I-labeled hypoiodite. A second portion was iodinated with 131I-labeled hypoiodite in the presence of hapten to protect the site from iodination. The protected and unprotected preparations were mixed, digested with pepsin and the iodinated peptides separated. The ratio of 125I/131I relative to that of the unfractionated digest was determined for the peptides; a high ratio was taken to indicate that the peptide was derived from the antibody site. Two high-ratio peptides, each in a yield of about 30% of the total antibody site initially present and probably from different populations of the anti-3P antibody, had the sequences Val-Ser-MIT (monoiodotyrosine) and MIT-Ser-Leu. The two peptides were shown to be derived from the heavy chain of the antibody and probably from the variable portion. A third high-ratio peptide, present in a much smaller amount (5% yield), was from the light chain fraction.
Antibody Response in Genetically Thymus-Less Nude Mice Injected with Normal Thymus Cells
Authors: Kindred Berenice
Published: December 2022

Nude (nu/nu) mice, which lack the thymus, failed to produce an immune response to various antigens. When such animals were injected intravenously with normal thymus cells, lasting immune responsiveness was restored only if the cells came from animals “related” to the nude recipients. The required relationship was probably H-2 compatibility. An allotype marker was used to determine the specificity of the antibody-producing cells. Even after a period of 12 weeks the antibody produced was entirely of host cell type. No evidence that thymus cells may develop into antibody-producing cells was found.
Effect of Antigen Variation on Production of Antibody in Canine Tracheal Secretions
Authors: Lieberman Phillip , Patterson Roy , Petersen Verlyn , Chakrin Lawrence W. , Wardell Joe R. , Ricks John
Published: December 2022

Mongrel dogs with tracheal pouches were immunized by pouch injection with bovine serum albumin (BSA), keyhole limpit hemocyamin (KLH) and sheep red blood cells (SRBC). Antibody responses were assayed in tracheal secretions and in serum. Antibody apparently gained access to tracheal secretions by two means. Antibodies to KLH and BSA in tracheal secretions appeared to have been produced systemically and transported to the pouch and into the secretions. In contrast, antibodies to SRBC were apparently manufactured locally as well as systemically, as shown by the presence of an IgA anti-SRBC that was detected in tracheal secretions but not in sera.
Immune Fever in the Rabbit: Responses of the Hematologic and Complement Systems
Authors: Mickenberg Ira D. , Snyderman Ralph , Root Richard K. , Mergenhagen Stephen E. , Wolff Sheldon M.
Published: December 2022

Non-pyrogenic human serum albumin (HSA) administered to specifically immunized rabbits or given as HSA-anti HSA complexes to normal recipients induced fever, leukopenia, thrombocytopenia and consumption of complement. The pattern of these responses was distinctive from those induced by endotoxin of comparable pyrogenicity. Whereas the degree of in vitro complement consumption related directly to the antigen-antibody ratio, that observed in vivo after complex administration was more dependent upon the quantity of complexes given and their duration in the circulation. The rate of clearance of complexes from the circulation was independent of the degree of complement consumption but dependent upon the antigen-antibody ratio of the complexes. In general, no correlation was found between the extent of in vivo complement consumption and the degree or duration of fever, leukopenia or thrombocytopenia, although at low dosages the degree of complement consumption paralleled febrile responses.
Schistosomal Egg Antigen-Induced Lymphocyte Blastogenesis in Experimental Murine Schistosoma mansoni Infection
Authors: Colley Daniel G.
Published: December 2022

Cultures of lymph node cells from mice infected with Schistosoma mansoni underwent a vigorous blastogenic reaction in response to a soluble antigen preparation derived from schistosomal eggs. Lymph node cell cultures from normal mice failed to respond to this antigen. Dose response studies demonstrated that as little as 1 µg protein of the antigen preparation would induce marked specific tritiated thymidine incorporation in cultures of infected mouse lymphocytes. The kinetics of this response are comparable to those seen in other instances of antigen-induced blastogenesis. Neither the response to phytohemagglutinin nor that to pokeweed mitogen was altered by schistosomal infection.
Enhancement of Hemolytic Complement Activity by Treatment of Human Serum with Iodine
Authors: Polley Margaret J.
Published: December 2022

A method has been described for enhancing the biologic activity of human complement by treatment of whole human serum with a critical concentration of iodine. The effect was due to oxidation of the second component of the complement system (C2) contained in the serum. This chemical modification increased the activity of this component to the point at which it no longer limited the complement reaction sequence.
Relation of the Human Antibody Response to Flagellin to Gm Genotype
Authors: Wells J. Vivian , Fudenberg H. Hugh , MacKay Ian R.
Published: December 2022

The incidence and distribution of the Gm allotypic markers Gm (a), (x), (g), (b) and (f) were studied in 113 Caucasian subjects who were classified on the basis of the serum titer of “natural” anti-flagellin antibodies and their response to injection with 5µg flagellin from Salmonella adelaide. There were 59 subjects with a low titer (< 100) of “natural” anti-flagellin antibodies (LNA) and of these, 43 had a low primary response (LPR, peak titer < 100) to injected flagellin; these 59 subjects were classified as “low responders.” There were 18 subjects with a high titer of “natural” antibodies (HNA, titer τ; 600) and a separate group of 36 with a high primary response (HPR, titer τ; 10,000); these 54 subjects were classified as “high responders.”.

As expected in Caucasians, all subjects positive for Gm (a) were also positive for Gm (g) (64.6%) and all subjects positive for Gm (b) were also positive for Gm (f) (89.4%). The number of Gm (a,g) subjects was significantly higher in group HNA than in group LNA (P < 0.05) and significantly higher in “high responders” than in “low responders” (P < 0.05). Otherwise stated, the absence of the Gma,g gene group was associated with a lower response to flagellin. The results indicate a significant association in these subjects between the magnitude of the immune response to flagellin (indicated by antibody titers) and the Gm genotype.
The Importance of θ and Ig Bearing Cells in the Immune Response to Various Antigens
Authors: Takahashi T. , Mond J. J. , Carswell E. A. , Thorbecke G. J.
Published: December 2022

Transfers of normal and/or immune BALB/c spleen cells into x-irradiated syngeneic recipients were performed in order to characterize further the nature of memory cells. Incubation of the cells with antisera to either the θ-antigen or to mouse immunoglobulin in the presence of C was used to remove selectively T-cells (θ-bearing) or B-cells (Ig-bearing).

Primary and secondary responses to Brucella abortus (BA) were reduced by incubation with anti-Ig but were unaffected by removal of T-cells, suggesting the thymus-independence of this response. Responses to sheep erythrocytes (SE) were always greatly inhibited after incubation of the cells with anti-θ, while being reduced to a variable extent by anti-Ig. With DNP-hemocyanin as the antigen, primary and early memory responses were unaffected by anti-θ, whereas the late memory response to this antigen was greatly reduced by such treatment.

Addition of equal numbers of untreated normal spleen cells to antisera treated immune spleen cells resulted in minimal reconstitution of the memory responses. Such reconstitution was only obtained by recombination of anti-θ and anti-Ig treated immune spleen cells. It is concluded that immunologic memory can be expressed by both the B- and the T-lymphoid cell lines in the mouse.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Effect of anti-γ chain and anti-µ chain on secondary antibody response of rabbit spleen cells was studied in vitro. Spleen cells from rabbits primed with dinitrophenylbovine γ globulin (DNP-BGG) were stimulated in vitro with the antigen either in the presence or absence of anti-heavy chain antibody. After 24 hr of incubation, the cells were washed and cultured in the absence of the antigen. Antibody response was evaluated by enumerating IgG and IgM antibody producing cells and by measuring anti-DNP antibodies in culture media. It was found that anti-γ chain suppressed both IgG and IgM antibody response, while anti-µ chain suppressed only IgM antibody response. When the primed cells were cultured without antigen stimulation, IgM antibody producing cells increased after 3 days' culture. The response was also suppressed by treatment of the cells by either anti-γ or anti-µ chain. The treatment with anti-heavy chains did not inhibit secretion of antibodies from antibody producing cells. It appears that anti-heavy chain suppressed differentiation of precursor cells to antibody-producing cells.
Cell Surface Associated Gamma Globulins in Lymphocytes
Authors: Lee S.-T. , Paraskevas F. , Israels L. G.
Published: December 2022

The reverse immune cytoadherence test (RICA) was modified to enable the detection of two different classes of surface associated γ globulins on mouse spleen lymphocytes (mixed RICA). This was achieved by the use of two 5S hybrid antibodies each carrying one anti-γ globulin site highly specific for one class of γ globulin. Two different indicator cells were used to demonstrate the binding of both hybrid antibodies on the same lymphocyte, thus forming a mixed rosette. With the mixed RICA we were able to determine that per 1000 spleen cells approximately 200 lymphocytes carry both major mouse γ globulin classes, γG (7Sγ2a) and γF (7Sγ1) on their surface and 100 lymphocytes carry only γF globulin. Few cells were detected carrying only γG globulin. If the surface γ globulins indicate the potential of the cell to synthesize that γ globulin our results indicate that the majority of mouse spleen “resting” lymphocytes are pluripotential. The co-existence of unipotential cells (carrying only γF globulin) may result from previous antigenic stimulation.
Latent Measles Virus Infection of the Hamster Central Nervous System
Authors: Wear Douglas J. , Rapp Fred
Published: December 2022

Human measles virus adapted to the hamster brain replicated to high titers and initiated neural signs of encephalitis when incoulated intracerebrally into unprotected newborn hamsters. However, the presence of high levels of maternally acquired measles neutralizing antibody blocked the onset of acute encephalitis without interrupting intracellular virus replication. the measles infection remained latent until cyclophosphamide was introduced. A large percentage of the immunosuppressed hamsters developed chronic persistent myoclonic tremors of various muscular areas. Virus was isolated from these animals as long as 90 days post-infection. Thus, the newborn hamster is a useful model for the study of latent central nervous system infection by measles virus.