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E F Osserman

Publications and source records attributed to E F Osserman.

At least 37 records · Page 2Linked to original sources
PubMed1982

Quantitative recovery, selective removal and one-step purification of human parotid and leukemic lysozymes by immunoadsorption.

Immunoadsorption affinity chromatography was used to isolate and purify human lysozyme. The immunoadsorbent was prepared by coupling sheep anti-(human leukemic lysozyme) IgG to epoxy-activated Sepharose 6B. Lyophilized parotid saliva (21) was resuspended in distilled water (325 ml, 50 mg/ml, w/v) and applied to a column which had a capacity to bind 4.25 mg human enzyme. Non-adsorbed material did not contain lysozyme, as determined by enzymatic and immunological analyses. All lysozyme activity present in the applied sample (1.97 mg) bound to and was desorbed from the column by elution with 0.2 M sodium acetate HCl buffer, pH 1.8. The isolated material was homogeneous as determined by cationic and sodium dodecyl sulfate/polyacrylamide gel electrophoresis, ultracentrifugation, amino acid and amino-terminal analyses, and immunoelectrophoretic analysis. The one-step purification procedure yielded a 1370-fold increase in specific activity. Human lysozyme was also selectively purified by this method from an ammonium sulfate precipitate of the urine of a patient with chronic monocytic leukemia. Amino acid and polyacrylamide gel electrophoretic analyses indicated that the purified enzyme was identical to human lysozyme isolated from leukemic urine by classical biochemical techniques.

Chromatography, Affinity↗
PubMed1982

Identical twin marrow transplantation in multiple myeloma.

We performed the first successful syngeneic bone marrow transplantation (BM Txp) in a patient with multiple myeloma. The patient and his normal identical twin are 50-year-old physicians. Prior to BM Txp, a partial remission was achieved with 1 year of continuous low dosage melphalan and prednisone therapy. Immediately before BM Txp, high dose cyclophosphamide and total body irradiation were administered in an attempt to eradicate the residual tumor. For 17 months after BM Txp, the patient was asymptomatic and hematologically normal although a low concentration of serum monoclonal IgGK persisted. In the 18th month, recurrence of bone pain and increase in the monoclonal IgG signalled exacerbation of the disease. Chemotherapy was resumed and again produced objective and subjective evidence of response. This study demonstrates the feasibility and potential usefulness of syngeneic BM Txp in myeloma.

Blood Grouping and Crossmatching↗
PubMed1982

Spontaneous in vitro differentiation of antigen-specific lymphocytes from a patient with immunoglobulin M gammopathy.

Recently we have identified two monoclonal immunoglobulin M (IgM) proteins that bind Klebsiella polysaccharides. The lymphocytes of one of these patients (M.A.Y.) were available for study. A substantial proportion of the B lymphocytes isolated from this patient's peripheral blood also bound Klebsiella polysaccharides with a pattern of specificity identical to that of the monoclonal IgM, and reacted with an anti-idiotypic antiserum directed against this IgM. Stripping the surface immunoglobulin from these lymphocytes eliminated this reactivity. Although no plasma cells were detected in the freshly isolated peripheral blood lymphocytes of this patient, plasma cells binding Klebsiella polysaccharide appeared after 7 d of in vitro culture. This occurred regardless of whether the cultures were supplemented with autologous plasma, normal human plasma, or fetal calf serum. Pokeweed mitogen neither stimulated nor inhibited the in vitro differentiation of the monoclonal B lymphocytes into plasma cells. This differentiation was, however, abrogated by F(ab')2 fragments of anti-human IgM and by anti-idiotypic antibodies, as well as by the Klebsiella polysaccharide with which the monoclonal IgM reacted.

Aged↗
PubMed1982

Lysozyme stimulates lymphocyte proliferation in monocyte-depleted mixed lymphocyte cultures.

Human LZM was found to enhance 3H-thy uptake and lymphoblast transformation in monocyte-depleted MLC. The effect was maximal (up to sixfold increase) in "low level" (less than 10,000 cpm) MLC. Maximal MLC enhancement was obtained with 250 micrograms/ml LZM; higher LZM concentrations appeared to be inhibitory. LZM enhancement of MLC could not be demonstrated in the presence of monocytes. LZM did not enhance lymphocyte responses to SKSD. Several observations support the possibility that LZM enhancement of MLC is due to LZM augmentation of stimulator cell antigenicity. LZM pretreatment of stimulator but not responder lymphocytes enhanced MLC. This enhancement was blocked by the LZM inhibitor tri-GlcNAc. LZM enhancement of MLC was maximum when the antigenic stimulus was minimized by either reducing the number of stimulator cells or utilizing DRw-matched donors in MLC. These data suggest that (1) LZM may enhance lymphocyte proliferative responses to allogeneic stimuli by altering stimulator cells' antigenicity and (2) monocyte-macrophage enhancement of lymphocyte proliferation in MLC may be partially mediated by LZM. (J Lab Clin Med 99:370, 1982.)

B-Lymphocytes↗
PubMed1982

A monoclonal IgM lambda macroglobulin with specificity for lacto-N-tetraose in a patient with bronchogenic carcinoma.

The serum of a patient with bronchogenic carcinoma was found to contain a monoclonal IgM lambda (IgMwoo) that precipitated with a precursor blood group glycoprotein containing I and i determinants. IgMwoo did not agglutinate O cells in the cold or at room temperature, and by quantitative precipitin and precipitin inhibition assay its specificity was shown not to be to the I and i determinants. IgMwoo reacted best with lacto-N-tetraose, DGal beta 1 leads to 3DGlcNAc beta 1 leads to 3DGal beta 1 leads to 4DGlc, and was specific for the non-I or non-i determinant dGal beta 1 leads to 3DGlcNAc beta 1 leads to 3DGal-moiety present as a distinct chain on precursor glycoproteins containing I and i determinants. Human ovarian cyst blood group A and B glycoproteins were inactive, but removal of the outer tier of sugars involved in A, B, and H specificity exposed this non-I or non-i determinant as well as the determinant reacting with anti-I Ma. IgMwoo was neither a cold agglutinin nor a cryoglobulin. It precipitated with precursor blood group glycoproteins somewhat less at 37 degrees C than at 0 degrees C, the differences being ascribable to solubility.

ABO Blood-Group System↗
PubMed1981

Further studies of the riboflavin-binding immunoglobulin IgGGar. Resolution into fractions of different riboflavin content and aspects of reassembly.

A previously described human immunoglobulin with unusual flavin-binding activity, IgGGar [Farhangi, M., & Osserman, E. F. (1976) N. Engl. J. Med. 294, 177], is further characterized. The protein can be fractionated into two subpopulations, one of which is nearly completely saturated with riboflavin and one in which the binding sites are largely vacant. Possible differences between these fractions and/or their binding sites are explored. While electrophoretically distinct, the IgGGar-riboflavin complexes possess a basic similarity in the binding sites of both fractions as evidenced by spectroscopic examination. However, an important difference exists in that added riboflavin equilibrates reversibly with the vacant sites of native IgGGar, while the riboflavin in the occupied sites is essentially irreversibly bound. The tight association may be due to an in vivo combination of riboflavin with protein of different conformation than occurs in vitro, such as an incompletely assembled or folded tetramer. Accordingly, in vitro renaturation was examined. Studies of renaturation revealed that the reduced interchain disulfides within a tetramer reoxidize smoothly, although inter-heavy-chain bonds form less readily than inter-heavy-light-chain disulfides. Renaturation of IgGGar, unlike previously studied IgG molecules, does not proceed under conditions in which the protein structure had previously been significantly disrupted. The assembly defect is localized in the inability of the denatured heavy chain to refold into a stable species capable of combining with the light chain.

Binding Sites↗
PubMed1981

Localization of the carbohydrate units in a human immunoglobulin light chain, protein Sm lambda.

The carbohydrate structure and complete amino acid sequence of a human lambda-type immunoglobulin light chain, protein Sm lambda has been determined. The protein was isolated from the urine of a patient with a plasma cell dyscrasia resembling gamma-heavy-chain disease. 13 tryptic peptides covering the entire polypeptide chain of 135 residues were isolated from the aminoethylated protein, and 15 chymotryptic peptides, accounting for 131 residues, were recovered from the carboxymethylated protein. The sequence of 18 of these peptides was partially or completely determined by the Edman-dansyl technique or C-terminal analysis, permitting the establishment of the complete primary structure of the polypeptide chain. The sequences established that this light chain possessed an intramolecular deletion of 81 amino acid residues. The N-terminal 30 residues showed considerable homology with other lambda chains of subgroup II. The defect began at position 31, in the first hypervariable region, and encompassed the remainder of the variable region through position 109. The constant region was fully intact and normal synthesis recommenced with a glutaminyl residue at position 110, the first residue of the constant region. This light chain contained carbohydrate in the hypervariable region just preceding the deletion. The precise number and locations of the oligosaccharide chains were established by amino acid sequence analysis of glycopeptides isolated from proteolytic hydrolysates by chromatography on Bio-Gel P-6 columns. These studies showed that protein Sm lambda contains one N-glycosidically-linked chain attached to asparagine-25 and one O-glycosidically-linked oligosaccharide chain attached to serine-21. The structures of the oligosaccharide chains were determined by methylation analysis, gas chromatography and hydrolysis with specific glycosidases. The structure of the N-glycosidically-linked chain was NeuAc(alpha 2 leads to 6)Gal(beta 1 leads to 4)GlcNAc(beta 1 leads to 2)Man(alpha 1 leads to 6)[NeuAc(alpha 2 leads to 6)Gal(beta 1 leads to 4)GlcNAc(beta 1 leads to 2)Man(alpha 1 leads to 3)]Man(beta 1 leads to 4)GlcNAc(beta 1 leads to 4)[Fuc alpha 1 leads to 6]GlcNAc leads to Asn. The second O-glycosidically-linked chain was a disialylated tetrasaccharide with the structure, Neu(alpha 2 leads to 3)Gal(beta 1 leads to 3)[NeuAc(alpha 2 leads to 6)GalNAc leads to Ser. This mucin-type disialylated tetrasaccharide in close proximity to N-asparagine-linked chains has not been previously observed in the oligosaccharide chains of immunoglobulins.

Amino Acid Sequence↗
PubMed1980

Human monoclonal macroglobulins with specificity for Klebsiella K polysaccharides that contain 3,4-pyruvylated-D-galactose and 4,6-pyruvylated-D-galactose.

Two human IgM myeloma proteins, IgMWEA and IgMMAY, were found to react with agar and Klebsiella polysaccharides that contain pyruvylated D-galactose (DGal). Quantitative precipitin data and precipitin inhibition studies with methyl alpha- and beta-glycosides of 4,6-pyruvylated-D-galactose showed their combining sites to be different, although each was directed against the pyruvylated-D-Gal, one reacting most specifically with Klebsiella polysaccharides with terminal nonreducing beta-linked 2,4 pyruvylated-D-Gal, whereas the other reacted equally well with Klebsiella polysaccharides that contain 3,4 beta-linked and 4,6 alpha-linked terminal nonreducing pyruvylated-DGal. Inhibition studies showed that both sites are directed toward one of the two space isomers of 3,4- or 4,6-pyruvylated DGal, the form in which the methyl group of the pyruvate is equatorial, or endo, and its carboxyl group axial, or exo, to the plane of the acetal ring. Coprecipitation studies showed the combining site of IgMWEA to be located on an (Fab')2 fragment and not on the (Fc)5mu fragment. The monoclonal peak in the serum of IgMMAY was specifically precipitated by Klebsiella polysaccharide. Myeloma proteins with specificities of this type may occur with reasonable frequency in humans and may be a consequence of clonal expansion from inapparent infection, carrier states, or disease produced by various Klebsiella organisms.

Adult↗
PubMed1980

Effects of dichloromethylene diphosphonate on skeletal mobilization of calcium in multiple myeloma.

Dichloromethylene diphosphonate (Cl2MDP), an inhibitor of oestoclast activity, was evaluated for its ability to decrease the excessive mobilization of skeletal calcium that complicates multiple myeloma. Ten patients with active myeloma, wide-spread bone disease, and hypercalciuria were studied in a double-blind, placebo-controlled, crossover-designed trial in which they took Cl2MDP for eight weeks and placebos for eight weeks. Two patients died during the placebo phase; of eight patients who received Cl2MDP, seven had rapid, sustained, and highly significant (P less than 0.001) decreases in urinary excretion of calcium. Six also had significant decreases in hydroxyproline excretion, and five reported lessening of skeletal pain. On patient did not respond. Although the patients received concurrent chemotherapy during the study, concentrations of myeloma proteins actually increased or decreased only slightly, indicating the declines in hypercalciuria resulted from Cl2MDP and not from improvement in the underlying disease. We conclude that Cl2MDP is a potentially useful inhibitor of osteoclast-mediated bone erosion in multiple myeloma.

Adult↗
PubMed1979

Modulation of neutrophil function by lysozyme. Potential negative feedback system of inflammation.

Host responses to infectious organisms should be modulated so that tissue-damaging products of inflammatory cells do not produce excessive destruction of normal tissue. Lysozyme, which is continuously secreted by monocytes, which, in turn, migrate relatively late to inflammatory areas, was found to significantly dampen several responses of neutrophils to inflammatory stimulants. Thus, human lysozyme obtained and purified from the urine of patients with monocytic leukemia (but not its structurally similar and comparably cationic analogue, eggwhite lysozyme) depresses chemotaxis of normal neutrophils to activated complement, bacterial supernate, and N-formylmethionyl-phenylalanine. In addition, human (but not eggwhite) lysozyme depresses oxidative metabolism (hexose monophosphate shunt activity) and superoxide generation of neutrophils. The specificity of the suppressive effects was indicated by inhibition studies with rabbit antihuman lysozyme antibody, and with the trisaccharide of N-acetylglucosamine, a specific inhibitor of lysozyme. The results suggest that lysozyme, a product of inflammatory cells themselves, may function in a negative feedback system to modulate the inflammatory response.

Cell Migration Inhibition↗
PubMed1978

Computer-based case tracing (COMTRAC).

To assist clinical assessment and decision making, a computer-based system has been developed to organize key data of individual medical records and display these data in a standard graphic format for each disease. A program for multiple myeloma has been completed and 150 case records entered. The data accessed by computer-based case tracing on all cases of a particular disease are available for cumulative and comparative analyses of different therapeutic regimens.

Data Display↗
PubMed1978

Serum lysozyme, serum proteins, and immunoglobulin determinations in nonspecific inflammatory bowel disease.

The serum levels of lysozyme, serum electrophoresis, and serum immunoglobulins were determined prospectively in 101 patients with ulcerative colitis, ulcerative proctitis, Crohn's disease, or nonclassifiable nonspecific inflammatory bowel disease. Although the mean serum lysozyme concentration of patients with Crohn's disease (10.5 +/- 6.8 microgram/ml) and ulcerative colitis (9.6 +/- 4.1 microgram/ml) performed by a standardized lysoplate method was significantly greater than normal controls (6.0 +/- 1.5 microgram/ml), the results did not correlate with the diagnosis nor with the degree of disease activity. Individually separated protein fractions and serum immunoglobulins also did not correlate with the serum lysozyme levels. This study indicates that measurement of the level of serum lysozyme in individual patients is not helpful in determining the cause or degree of activity of nonspecific inflammatory bowel disease.

Adult↗
PubMed1977

Primary structure of rat lysozyme.

For evolutionary reasons, we determined the primary structure of rat lysozyme. The chymotryptic peptides from the reduced and carboxymethylated protein were sequenced and aligned by homology with the sequence of human lysozyme. Overlaps were confirmed by partial structures of tryptic peptides and an automatic sequencer run on the whole protein. By comparing this lysozyme sequence with those of human and baboon and taking into account paleontological estimates of the times of divergence of these species from one another, an approximate estimate of the average rate of lysozyme evolution was made. This rate is not significantly different from the average rate of lactalbumin evolution in mammals--a finding which is at variance with Dickerson's [Dickerson, R.E. (1971), J. Mol. Evol. 1, 26] and Dayhoff's [Dayhoff, m.o., ed. (1972), Atlas of Protein Structure and Sequence, Vol. 5, Silver Spring, Md., The National Biomedical Research Foundation] conclusion that lactalbumin evolution has been faster than lysozyme evolution. Our finding raises the possibility that the gene duplication event responsible for the origin of lactalbumin from lysozyme was more ancient than is generally supposed. Furthermore, from comparison of the rates of lysozyme evolution in rodents and primates, it is suggested that generation time is not a key factor in lysozyme evolution.

Amino Acid Sequence↗
PubMed1976

Myeloma with xanthoderma due to an IgG lambdamonoclonal anti-flavin antibody.

When yellow skin and yellow hair developed in an elderly patient with multiple myeloma, we ruled out the usual causes of such pigmentation but identified a monoclonal IgGlambda (lgGGar) with anti-flavin antibody activity. Purified IgGGar was bright yellow, and the acid-dissociated chromophore was identified as riboflavin by chromatography and absorption spectroscopy. Native IgGGar contained 1.45 moles of flavin per mole of IgG, and increased to 2 moles with addition of riboflavin to saturation. The flavin was localized to the Fab fragment and was bound to IgGGar with high affinity. IgGGar showed strongest affinities for riboflavin, flavin mononucleotide and flavin adenine dinucleotide, and lower affinities for dinitrophenyl derivatives and naphthoquinone. The demonstration of hapten bound to the circulating monoclonal immunoglobulin in this case suggests the possibility of bound but colorless haptens on other myeloma proteins as well as on normal immunoglobulins.

Aged↗