Reaction Of Partial Identity Research Articles

Örjan Ouchterlony and the antigen-antibody double diffusion-in-gel: a survey.

The Swedish scientist Örjan Ouchterlony published four ground-breaking papers 1948-1966 in Acta Pathol Microbiol Scand where he described a new method of antigen-antibody reactions in gel. He described and defined the 'reaction of identity' and 'reaction of partial identity' when he used related antigens and 'reaction of non-identity' when he used non-related antigens. His results inspired scientists in other countries to further develop and modify the 'Ouchterlony method' which became useful for both scientific and clinical purposes. This survey describes how the methods were discovered and how they became modified and improved and how they were used, but also underlines that the original Ouchterlony method is still used.

Alternanthera mosaic virus Found in Scutellaria, Crossandra, and Portulaca spp. in Florida.

In the fall of 1998, the Division of Plant Industry (DPI) received vegetative propagations of Scutellaria longifolia (skullcap) with symptoms of foliar mosaic, chlorotic/necrotic ringspots, and wavy line patterns from a nursery in Manatee County. Flexuous particles approximately 500 nm long were found with electron microscopy. The plants tested positive for Papaya mosaic virus (PaMV) in an enzyme-linked immunosorbent assay (ELISA) test with antiserum to PaMV (Agdia, Elkhart, IN). However, in immunodiffusion tests (antiserum from D. Purcifull, University of Florida), this virus gave a reaction of partial identity indicating it was related but not identical to PaMV (1). The original infected plants were kept in a greenhouse. In January 2005, a specimen of Crossandra infundibuliformis (firecracker plant) with mosaic symptoms was submitted to the DPI from a nursery in Alachua County. Inclusions found with light microscopy and particles found with electron microscopy indicated that this plant was infected with a potexvirus. This was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) with primers designed to detect members of the virus family Potexviridae (3). These plants reacted positive to PaMV antiserum in ELISA and gave a reaction of partial identity to PaMV in immunodiffusion. A specimen of Portulaca grandiflora (moss rose) with distorted leaves found at a local retail store was also tested and gave the same results. Leaves from each of the three plant species were rubbed onto a set of indicator plants using Carborundum and potassium phosphate buffer. Total RNA was extracted from symptomatic indicator plants of Nicotiana benthamiana. RT-PCR (3) was performed, and PCR products were sequenced directly. Sequences of approximately 700 bp were obtained for all three plant species and showed 98% identity with each other. BLAST search results showed that these sequences were 93% identical to an Alternanthera mosaic virus (AltMV) sequence at the nucleotide level but only 76% identical to PaMV. The amino acid sequences were 98 and 82% identical to AltMV and PaMV, respectively. The PCR products of the virus from Scutellaria sp. were cloned, resequenced, and the sequence was entered into the GenBank (Accession No. DQ393785). The bioassay results matched those found for AltMV in Australia (2) and the northeastern United States (4), except that the Florida viruses infected Datura stramonium and Digitalis purpurea (foxglove). The virus associated with the symptoms of these three plants appears to be AltMV and not PaMV. AltMV has been found in ornamental plants in Australia, Italy, and the United States (Pennsylvania, Maryland, and now Florida). Since this virus is known to infect several plants asymptomatically and can be easily confused with PaMV serologically, it is likely that the distribution of this virus is much wider than is known at this time.

Heterogeneity observed in the components of hemolysin BL, an enterotoxin produced by Bacillus cereus

Hemolysin BL (HBL), a diarrheal enterotoxin originally isolated from Bacillus cereus strain F837/76, is composed of three antigenically distinct proteins designated B, L 1, and L 2. All three components are required for biological activity. Here, we report antigenic and physical variations in HBL components produced by other B. cereus isolates. Reactions of partial identity were observed in double gel immunodiffusion assays using antibodies to highly purified B, L 1, and L 2 components of F837/76 and culture supernatants of strains F837/76 and S1C. Western blot analysis showed that F837/76 produced one 38-kDa B protein, one 38-kDa L 1, and one 43-kDa L 2 protein. In strain S1C, two B (38 and 42 kDa), two L 1 (38 and 41 kDa), and one L 2 (43 kDa) proteins were detected. Further Western blot analysis of 127 B. cereus isolates showed that 90 produced one or more of the three HBL components. Approximately half of these 90 isolates (43/90; 48%) produced protein profiles which differed from that of F837/76. A total of four B, two L 1, and three L 2 component profiles with proteins of different sizes were observed. Individual strains produced various combinations of single or multiple bands of each component. In addition, some strains produced only one or two of the three HBL components. The public health significance of these strains is unknown, as all three components are required for biological activity. The data presented here demonstrates a high degree of heterogeneity in HBL and provide the basis for further studies to characterize the variations in HBL and to determine the role of the variant components in pathogenicity.

Immunoelectrophoretic Characterization of Insulin Monomers and Polymers

Insulin monomers and polymers were analysed by quantitative immunoelectrophoretic procedures. The Zn-insulin hexamer dissociated reversibly by dialysis against the Zn-free electrophoresis buffer. The Zn-insulin polymers showed precipitin reactions of partial identity. Monomeric salt-free insulin migrated as soluble immune complexes in the antiserum gel. The insulin monomer did not absorb the precipitating antibodies against the Zn-insulin polymers. Thus the polymer structure creates antigenic epitopes absent from the insulin monomer. As insulin is probably released from the beta cells in the relatively stable form of Zn-insulin hexamers, selective monomer assays might underestimate the total content of immunoreactive insulin in the biological fluids. Electroimmunoassay of Zn-insulin immunoreactive antigens in human urine defines a normal reference range of 10-25 ng/ml.

Purification and characterization of two protein antigens from the heterogeneous BCG85 complex in Mycobacterium bovis BCG.

The heterogeneous BCG85 complex is a major component of BCG culture fluid. BCG85A and BCG85B were purified by combining ammonium sulphate precipitation with chromatography on hydroxyapatite, DEAE-Sephacel and phenyl-Sepharose columns. Twenty percent of BCG85B was recovered. The chromatographic separation procedures were monitored by fused rocket immunoelectrophoresis. The BCG85 complex was found to consist of three antigens, which were heterogenous with regard to electrophoretic mobility, molecular weight (MW), hydrophobic and immunological properties. They were designated A, B and C in increasing order, according to their electrophoretic mobilities. Thus BCG85A had the lowest electrophoretic mobility, BCG85C the highest. The MW of BCG85A was found to be 31,000, while BCG85B had a slightly lower MW, 29,000, as determined by SDS-PAGE. The antigenic relationship between the components was evaluated by crossed immunoelectrophoresis and double diffusion, and reactions of partial identity between the antigens were found. The BCG85 complex occurs in far lower concentration in sonicates of BCG than in culture fluid.

Variation of a 470 000 daltons antigen complex and catalase antigen in clinical isolates of Aspergillus fumigatus.

Antigens in ruptured mycelium of 18 Aspergillus strains including 14 clinical isolates of A. fumigatus were studied by immunoelectrophoresis. One antigenic component of molecular weight 470 000 previously characterized by hydrophobic interaction chromatography and gel filtration and a second component with catalase activity were detected in all A. fumigatus isolates but in varying quantities. The 470 000 antigen complex cross-reacted with antigens in A. flavus and A. nidulans but not in A. niger or A. terreus. A. fumigatus catalase antigen cross-reacted with catalase in A. flavus, A. nidulans and A. terreus, but not in A. niger. One A. fumigatus isolate produced two catalase antigens showing a reaction of partial identity. A. flavus also produced two catalase antigens, one of which was species-specific.

Immunochemical characterization of rough Brucella lipopolysaccharides.

Lipopolysaccharides (LPS) were extracted from rough strains of Brucella abortus and Brucella melitensis and from strains of the naturally occurring rough species Brucella ovis and Brucella canis. Brucella rough lipopolysaccharides (R-LPS) were readily distinguished from Brucella smooth lipopolysaccharides (S-LPS) and enterobacterial R-LPS, by their chemical, physical, and serological characteristics. B. ovis R-LPS was differentiated from B. abortus, B. melitensis, and B. canis R-LPS by its reaction of partial identity in immunodiffusion. Monospecific mouse sera against B. ovis R-LPS agglutinated only the homologous bacteria but not R cells of other species of Brucella. B. ovis R-LPS contained more 2-keto, 3-deoxyoctonate, and glucosamine as a percentage of dry weight than any other R-LPS tested. B. abortus R-LPS was identified by the absence of an unidentified sugar present in the other R-LPS molecules, and B. melitensis R-LPS could be differentiated from B. canis R-LPS by its higher content of fatty acids. In contrast to S-LPS, all of the R-LPS studied lacked quinovosamine. In electron micrographs, Brucella R-LPS had a granular appearance, in contrast to typical lamellar structures formed by Brucella S-LPS and Escherichia coli R-LPS.

Chromatographic fractionation of rhesus monkey ( Macaca mulatta) IgG subclasses using DEAE cellulose and protein A-Sepharose

Rhesus monkey serum was applied to a protein A-Sepharose column at pH 7.3 IgG was not detectable in the unbound effluent. Bound protein was eluted with a pH gradient of citrate-phosphate buffer. Two major elution peaks reproducibly resulted at pH 4.85–4.9 and pH 4.35–4.43, respectively, with only slight variability among individuals. Since the immunoelectrophoretic mobility of IgG in the peak at pH 4.85–4.9 was slower than that of the peak at pH 4.35–4.43, serum was first fractionated by ion exchange chromatography into 2 fractions: the first did not bind to DEAE cellulose in 0.01 M phosphate buffer pH 7.8, the second did bind and was eluted by an NaCl gradient. Each DEAE fraction was then further fractionated on protein A-Sepharose. IgG in the first DEAE fraction bound to protein A at pH 7.3 and eluted in a single peak at pH 4.72–5.0. IgG in the second DEAE fraction also bound to protein A, but eluted in 2 peaks at pH 4.65–5.1, and 4.25–4.6, respectively. All 3 IgG fractions eluted in the same position from an A5m column, had immunodiffusion reactions of identity with anti-human γ chain and were composed of similar heavy and light chains judged by SDS polyacrylamide gel electrophoresis. However, IgG eluting from protein A at the low pH (4.25–4.6) had a reaction of partial identity with other IgG fractions when tested by immunodiffusion against rabbit anti-rhesus γ chain, suggestive of an antigenically different subclass. Analysis of rabbit antisera prepared against the 3 IgG fractions confirmed the occurrence of at least 3 antigenically distinct rhesus monkey IgG subclasses. These 3 subclasses have been provisionally designated IgG I, IgG II and IgG III.

Chemical and serological properties of a cyanogen bromide peptide of southern bean mosaic virus protein.

Southern bean mosaic virus (SBMV) protein was cleaved with cyanogen bromide and a highly basic peptide, CB-1, was isolated by ion exclusion and ion-exchange chromatography. Twelve peptides were separated from a tryptic digest of CB-1 by ion-exchange chromatography and the composition of these peptides was similar to that of peptides released from EDTA-swollen virus particles by limited tryptic digestion. The composition and N-termini of the tryptic peptides indicated CB-1 was from the N-terminus of SBMV protein and contained 48 amino acid residues. The CB-1 peptide moved rapidly to the cathode in polyacrylamide gel electrophoresis at pH 3.9 and contained nine arginine residues, three lysine residues, and no acidic amino acid residues. It was shown to interact with purified viral RNA, sodium dextran sulfate, and calf thymus DNA. Antiserum to sodium dodecyl sulfate (SDS)-dissociated virus gave a reaction of partial identity between the CB-1 peptide and the SDS-dissociated virus in SDS gel diffusion tests. The CB-1 peptide did not react with antiserum to SDS-dissociated, trypsin-treated virus. Gel diffusion tests conducted in saline agar gels between trypsin-treated virus and SBMV, with SBMV antiserum, did not show differences in their serological properties. Antiserum to the CB-1 peptide conjugated to tomato bushy shunt virus reacted with SBMV but SBMV antiserum did not react with CB-1 or the CB-1-tomato bushy shunt virus conjugate.

Ocular antigens XI. Common antigenic determinants in the soluble retinal fractions of man, monkey, pig and calf

Common antigenicity between the soluble retinal fractions of man, monkey, pig and calf has been investigated by means of several immunological techniques. Lymphocytes of rabbits sensitized to human retinal antigens are equally stimulated by human, monkey and pig retina extracts. The results of immunoelectrophoresis point to the presence of immunologically identical antigens in human, monkey and pig retinas. In immunodiffusion, bovine and pig retinal antigens show only a reaction of partial identity with human retinal antigens. In immunoelectrofocusing, no precipitin lines are formed between basic retinal proteins of the tested species and rabbit anti-human retina antiserum. By means of the same technique reactive antigens with an IEP of 4·7 have been demonstrated in the soluble retina extracts of all investigated species. Both human and pig retina extracts contain a reactive antigen with an IEP of 5·3. They equally stimulated the lymphocytes of retinitis pigmentosa patients, which might be attributed to a sensitization of these patients to this antigen. In human, monkey as well as in bovine retinas an antigenic protein with an IEP of 5·9 was detected.

Evidence for the lipoprotein heterogeneity of human plasma high density lipoproteins isolated by three different procedures

AbstractQuantitative electroimmunoassays of apolipoproteins in ultracentrifugally isolated high density lipoproteins (HDL) of normolipidemic subjects showed that A‐I and A‐II are the major (80–85% of total HDL protein) and B, C‐III, E, D and F are the minor protein constituents of this density class. A comparison between the apolipoprotein composition of ultracentrifugally isolated HDL and heparin‐Mn++ supernates showed no significant difference in the levels of A‐I and C‐III. However, the concentration of ApoE in the heparin‐Mn++ supernates was almost twice as high as that in the ultracentrifugally isolated HDL; ApoB was only detectable in trace amounts in the heparin‐Mn++ supernates. To establish whether these apolipoproteins are parts of a single macromolecular complex or form separate, discrete lipoprotein particles, the high density lipoproteins were isolated by three different procedures including ultracentrifugation, heparin‐Mn++ precipitation and agarose column chromatography. The double diffusion analyses of each of these HDL preparations with antisera to A‐I, A‐II, ApoB, C‐III, ApoD, ApoE, and ApoF showed nonidentity reactions between each possible combination of these antisera. The only exception was a reaction of partial identity between antisera to A‐I and A‐II polypeptides indicating the occurrence of two types of lipoprotein particles, a major one (LP‐A) containing both polypeptides and a minor one (LP‐A‐I) containing A‐I as the sole protein constituent. These findings indicate that high density lipoproteins, regardless of the manner of isolation, do not consist of a single macromolecular complex, but represent a mixture of several, discrete lipoprotein families.

Experimental Allergic Uveitis. Isolation, Characterization, and Localization of a Soluble Uveitopathogenic Antigen from Bovine Retina

Abstract The soluble antigen involved in the pathogenesis of EAU was isolated from bovine retinal extract by precipitation with ammonium sulfate followed by molecular sieve and ion exchange chromatography. The purified fraction formed a double precipitin band by immunodiffusion and immunoelectrophoresis, and a doublet by analytical disc electrophoresis. By immunodiffusion one of these components formed a reaction of identity with guinea pig S antigen and the other a reaction of partial identity. Because the two components had the same m.w. and similar electric mobility, they are presumed to be minor structural variants of the same molecule. The purified antigen had a m.w. of approximately 102,000 by analytical ultracentrifugation, 50,500 by SDS electrophoresis, and 56,000 by gel filtration chromatography. Sedimentation velocity studies with increasing concentrations of antigen indicated self-association. An S20 value of 3.6 was obtained for the monomer, corresponding to an approximate m.w. of 50,000. The antigen was identified as a protein with a very small amount of lipid. Data are presented on the absorption spectrum, extinction coefficient, and amino acid composition. The molecule was lacking in cysteine and tryptophan, but contained a relatively high proportion of nonpolar amino acids corresponding with a lipophilic protein. It was localized by immunofluorescence to the area surrounding the entire photoreceptor cell in a pattern indicating association with the plasma membrane. The antigen was not rhodopsin, which has been implicated as a probable antigen in the pathogenesis of EAU, but has some properties resembling a recently described photoreceptor cell retinol-binding protein tentatively implicated in the transport of vitamin A from the circulation to the retina. Assay of the purified fraction for production of EAU in the guinea pig indicated it was highly pathogenic in the microgram range. In agreement with immunofluorescent findings, the photoreceptors were specifically destroyed after cellular infiltration of the uveal tract.

Are “hidden” genes for immunoglobulin allotypes of rabbit heavy and light chains expressed upon hyper‐immunization?

AbstractThe question of whether genes for some Ig‐allotypic forms may be “hidden” in the genome was investigated by hyperimmunization of 36 allotypically pedigreed rabbits with a vaccine of Micrococcus luteus (Micrococcus lysodeikticus). Since Strosberg et al. (J. Immunol 1974.113: 1313) have reported that such hyperimmunization elicited expression of a third “allele” at two different loci (a2 and b6 in a rabbit whose original phenotype was ala3b4b5), we wished to determine whether their immunization protocol would reproducibly elicit expression of “hidden” allotypes. None of the 36 immunized rabbits developed elevated expression of an unexpected allotype during the 7‐38 weeks of immunization and bleeding. However, we did observe elevated expression of unusual variable regions. In particular, a cross‐reaction between some VH region gene products produced by al rabbits, and the majority of VH products of a2 rabbits was detected with anti‐a2 antisera made in a3a3 but not in a1a1 or a1a3 rabbits. There is a small percentage of Ig molecules in a1a1 rabbits whose V regions contain an antigenic determinant also present on most or all a2 molecules of rabbits. A similar determinant is also found on Ig of related genera (Lepus and Sylvilagus) Expression of this population was elevated during immunization of an a1a3 rabbit with Micrococcus. In immunodiffusion, a reaction of partial identity was observed. In hemagglutination inhibition, the reaction of a3 anti‐a2 with homologous a2 was not inhibitable by the cross‐reacting component. Anti‐a2 antisera made in a3 rabbits could be depleted of cross‐reactive antibody by absorption with insolubilized sera from normal al rabbits but not by insolubilized a3 sera. Radioautography of isoelectric focusing plates overlayed with a3 anti‐a2 antibodies which had been affinity‐purified and radiolabeled, revealed distinct spectrotypes in sera from al rabbits that carried the cross‐reacting VH determinants.

Purification of Native Properdin by Reversed Affinity Chromatography and Its Activation by Proteolytic Enzymes

A highly purified preparation of human properdin (P) has been obtained in a simple two-step procedure utilizing reversed application of the technique of affinity chromatography. The method involved precipitation of properdin from human serum and subsequent passage through an immunoadsorbent column of Sepharose anti-RP globulin which bound the contaminating proteins. The immunochemical properties of the isolated properdin (P) was found to be different from those of activated properdin (P). P was shown to be a 6.1S, beta2 globulin with a mean subunit m.w. of 57,900. P on the other hand was a 5.1S protein with gamma2 mobility and a subunit m.w. of 53,000 daltons. Double diffusion analysis using anti-P revealed a reaction of identity between P and P. However, when the reaction was developed with anti-P, a reaction of partial identity was obtained and the precipitin line of P was seen to spur over the line developed with P. Mild treatment with plasmin or trypsin converted P to P. Unlike P, P was ineffective in triggering the activation of the Properdin System in RP unless trace amounts of zymosan were added. Under these conditions P was found to be converted to P. The results indicate that properdin is present in fresh serum in a precursor form and its activation to P involves a limited proteolytic cleavage of the molecule.

Immunochemical study of the peptidoglycan of gram-negative bacteria.

The specificity of antibodies directed against the peptidoglycan of gram-negative bacteria was studied. The peptidoglycans of Proteus vulgaris, Escherichia coli, Moraxella glucidolytica, Neisseria perflava, give identical precipitin reactions. By means of inhibition studies with various peptidoglycan subunits and synthetic peptides, it was shown that the antibodies are essentially directed against the peptide moiety of the peptidoglycan: L-Ala-D-Glu (L)-mesoA2pm-(L)-D-Ala, that the peptide reacts better with antibodies when it is not cross-linked, and that the C-terminal portion-meso-A2pm-D-Ala of the peptide is immunodominant. These results explain the immunological identity of the peptidoglycans of gram-negative bacteria, which possess the same peptide subunit. Only weak cross-reactivity was observed with the peptidoglycans of gram-positive bacteria (Streptococcus faecium, Micrococcus lysodeikticus, Corynebacterium poinsettiae) where meso-diaminopimelic acid is replaced by L-lysine or L-homoserine. However, the peptidoglycan of Bacillus megaterium which possesses the same peptide subunit as gram-negative bacteria, gives only a reaction of partial identity with these bacteria. This result suggests the presence on the peptidoglycan of gram-negative bacteria, of other undefined antigenic determinants.

Production of antiserum to human properdin and demonstration of antigenic differences between the native and activated protein.

Antisera were raised in rabbits to human properdin in the precursor form (P) and in the activated state (P). On Ouchterlony analysis using the anti-P, reactions of complete identity were obtained between P, P, and properdin in twofold concentrated serum (NHS). However, when anti-P was used, a reaction of identity was obtained only between P and NHS, and a reaction of partial identity was formed between P and P and between properdin in NHS and P, suggestive of the fact that certain antigenic determinants in P may be lacking in P. The results indicate that activation of precursor properdin may involve proteolytic cleavage and/or conformational alterations of the molecule.

Properties of Human Tissue Thromboplastins from Brain, Lung, Arteries, and Placenta

Properties of purified and delipidated human tissue thromboplastins (TTPs) from brain (BTTP), lung (LTTP), arteries (ATTP) and placenta (PTTP) were studied. The extinction coefficients were closely similar with a mean value of 8.3 +/- 0.01 (SD). The molecular weights were 200,000 +/- 3,000 (SD) BTTP), 90,000 +/- 2,000 (SD) (LTTP), 110,000 +/- 3,000 (SD) (ATTP) and 250,000 +/- 4,000 (SD) (PTTP). The maximum activity of each delipidated TTP after relipidation was obtained when phospholipid-delipidated TTP ratio was 2.0, and the maximum specific activity was 440 units per mg BTTP, 270 units per mg LTTP, 80 units per mg ATTP and 600 units per mg PTTP. Ouchterlony analysis with anti-delipidated PTTP antibody showed the reaction of partial identity between delipidated TTPs and the antibody. Studies of the reactivity of I-125-delipidated TTPs with the antibody gave the following average values for % bound I-125-TTPs in 2 hours: 3.6 +/- 0.2(SD) % (I-125-BTTP), 11.0 +/- 0.3 (SD) % (I-125-LTTP), 4.4 +/- 0.2 (SD) % (I-125-ATTP) and 13.7 +/- 0.3 (SD) % (I-125-PTTP). It was also found that the antibody markedly neutralized the coagulant activity of saline extracts of brain, lung and placenta. These results indicate that delipidated BTTP, LTTP, ATTP and PTTP are different while being similar in some aspects and that complete TTPs are not coated with lipids in a manner inaccessible to anti-delipidated PTTP antibody.

Detection of Membrane‐Bound Complement Components by Crossed Immunoelectrophoresis of Target Membranes Solubilized with Non‐Ionic Detergent

Sheep erythrocyte membranes lysed with human complement were solubilized in 1% Berol EMU‐043 (w/v) and subjected to crossed immunoelectrophoresis in the presence of this detergent. Using antibodies to human serum proteins, we found 5–6 protein precipitates whose presence was dependent on complement action. Among these, C3 and C4 components of complement were identified. Membrane‐bound C5 (C5b) did not precipitate in crossed immunoelectrophoresis, but its presence on target membranes was demonstrated by crossed‐line immunoelectrophoresis (in situ absorption) of specific anti‐CS antibodies. A reaction of partial identity between membrane‐bound C5 (CSb) and serum C5 was found. Our experimental approach allows immunoelectrophoretic analysis of complement components at the membrane level and should therefore prove useful for future investigations on the molecular structure of membrane‐bound complement.

Proteins in fluid from non-keratinizing jaw cysts. 3. Identification of individual proteins with particular reference to alpha- and beta-globulins, including fibrinogen.

Abstract. Fluids from 32 non‐keratinizing jaw cysts were examined by polyacrylamide gel disc electrophoresis, and by immunoelectrophoresis and double diffusion in agar against rabbit anti‐human serum and rabbit antisera monospecific for six different plasma proteins. Albumin, α1‐antitrypsin, α1‐acid glycoprotein (orosomucoid), transferrin, IgG, IgA, and IgM were invariably recognized. Most cyst fluids also reacted with antiserum to α1‐lipoprotein, α2‐macroglobulin, β1 C/β1A‐globulin (C3/C3c), and fibrinogen. Cyst fluid “fibrinogen” which showed somewhat slower electrophoretic migration than plasma fibrinogen, appeared as two precipitin bands or as one line with an apparently split cathodic end. Two cyst fluids exhibited one additional “fibrinogen” line located in the α1‐globulin region. C3, which was not detected in all cyst fluids, displayed an electrophoretic mobility between that of β1C‐ and β1A‐globulin of serum. In two cases an even faster C3 subtraction appeared. Double diffusion gave a “reaction of identity” between albumin and the α‐ and β‐globulins of cyst fluid and autologous scrum. Cystic “fibrinogen” and fibrinogen of human plasma gave a “reaction of partial identity”. These findings indicate that albumin and the main α‐ and A‐globulins of fluid from non‐keratinizing jaw cysts are derived from plasma and that breakdown products of fibrinogen or fibrin are also present. High levels of IgG and IgA, but decreased amounts of α2‐macroglobulin and C3 as compared to autologous serum are also suggested.

Immunologic relationships of antihemophilic factor of different species detected by specific human and rabbit antibodies.

SummaryThe plasmas of 14 nonhuman mammalian species corrected the specific clotting defect in human hemophilic plasma. The antihemophilic factor (AHF, Factor VIII) activity in all these plasmas was inhibited by human circulating anticoagulants directed against AHF and, except for rabbit AHF, by rabbit antiserum to purified normal human AHF, but human AHF was more readily inhibited than animal plasmas by both these agents. Porcine AHF was more resistant than bovine AHF to inhibition by all of 6 human circulating anticoagulants.In immuno-precipitation studies using rabbit antiserum, primate plasmas gave reactions of identity with human plasma, while non-primate mammalian plasmas gave reactions of partial identity. Plasma from a reptile and from two avian species contained no detectable antigen or clot-promoting properties related to human AHF.