David Catty

A quantitative study of the distribution of IgG sub-classes in a group of normal human sera.

The radial immunodiffusion method of Mancini has been applied to quantitative study of IgG subclasses in a normal population. The method was assessed in terms of both reproducibility and antiserum consumption. The distribution of IgG subclasses in a group of normal individuals was studied, and compared with their incidence in a series of monoclonal proteins investigated by previous workers and particularly with other quantitative studies on groups of normal individuals sera.

Defects in granulocyte function in various chromosome abnormalities (Down's-, Edwards'-, Cri-du-chat syndrome).

ZusammenfassungBei fünf Säuglingen mit Autosomenaberrationen, herabgesetzter Infektresistenz und intaktem humoralen und zellulären Immunapparat wurden die Granulocytenfunktionen Chemotaxis, Phagocytose, intrazelluläre Erregerabtötung und einige Stoffwechselvorgänge experimentell untersucht. Eine serumabhängige bzw. zelluläre Phagocytoseschwäche für Candida albicans bestand bei zwei Säuglingen mit Cri-du-chat-Syndrome und einem anderen mit Trisomie 18. Bei einem dieser Kinder bestand zusätzlich eine serumabhängige, verzögerte Erregerabtötung von Candida albicans sowie von Staphylococcus aureus, wobei die Serumspiegel der Opsonine IgG, IgM, CH 50 und C3 normal waren. Ein Säugling mit Trisomie 21 bot außer einem zellulären Chemotaxisdefekt ein verringertes zelluläres Abtötungsvermögen für Staphylococcus aureus sowie Escherichia coli im Eigen- und im Fremdserum. Die Phagocytose dieser Keime hingegen verlief ungestört.SummaryIn five infants with autosomal aberrations and diminished resistance to infection (in spite of intact humoral and cellular immune mechanisms) several granulocyte functions (chemotaxis, phagocytosis, intracellular killing and metabolism of killing) were measured. A serum-dependent or a cell-dependent disturbance of phagocytosis of Candida albicans was found in two infants with cat-cry syndrome and one with trisomy 18. In one of these children there was an additional serum dependent defect of the killing of Candida albicans and of Staphylococcus aureus, serum levels of opsonins (IgG, IgM, CH50 and C3) being within normal range. An infant with trisomy 21 showed, in addition to a cellular defect of chemotaxis, a reduced cellular ability of the killing of Staphylococcus aureus and of Escherichia coli in autologous and AB-pool-serum. Phagocytosis of these bacteria remained normal.

The application of cellulose carbonate to the preparation of water-insoluble immunoadsorbents used in the purification of antibodies to immunoglobulins

Abstract A new form of cellulose-derived immunoadsorbent has been developed which utilises cyclic trans-carbonate groups generated in the matrix to bring about covalent coupling of protein antigens. The production of ground material, the chemical coupling of immunoglobulin antigens and the characteristics of the immunoadsorbents for two systems, namely rabbit IgG and human IgM and their respective sheep antisera, is described. The adsorbents are relatively simple to prepare from the cyclic carbonate and in the systems described antigen coupling occurs under mild reaction conditions to produce insoluble reagents which are stable, highly specific to antibody and possess very high antibody binding capacities with respect to the amound of antigen coupled. Purified antibody can be eluted from the immunoadsorbent columns either by acid buffer or 3 M KCNS. Up to 86% of the adsorbed antibody can be recovered in purified form. This yield is sufficient to allow repeated use of columns for the large scale purification of antibodies, and far exceeds recovery of antiglobulins in most other published systems. Furthermore, since antibody binds to antigen in ratios which approach those in free solution, eluted antibodies differ little, if at all, from those in whole antiserum with respect to heterogeneity in specificity and affinity. In this latter property the cellulose carbonate-IgG adsorbent, in particular, was found to be more efficient than equivalent Sepharose cyclic imidocarbonate preparations.

The use of cellulose carbonate-based immunoadsorbents in the isolation of minor allotypic components of rabbit immunoglobulin populations.

Cellulose trans-2,3-carbonate has been used as a new insoluble matrix for the simple coupling of a1- and b4-positive rabbit immunoglobulin to make immunoadsorbents capable of purifying from serum, with great efficiency, alloantibodies to these allotypic determinants. The antibodies have themselves been conjugated to prepare specific antibody immunoadsorbents of high binding activity for their allotypic target molecules. With these anti-allotypic solid-phase reagents it has been possible to affinity purify a1- and b4-positive immunoglobulin molecules and to deplete serum immunoglobulin of these molecules to leave in the eluates only the allotypically uncontaminated minor immunoglobulin components which are a-negative or b-negative (lambda chain-bearing) molecules. lambda chain molecules were also purified in very small quantities by affinity chromatography on a sheep anti-rabbit lambda chain column. This method of purifying minor populations of rabbit immunoglobulin from normal serum by special immunoadsorbent applications offers new opportunities to study the products of rarely expressed immunoglobulin genes in normal rabbits.

Production of immunoadsorbents based on cellulose carbonates and their application to the isolation of specific immunoglobulin light chains

Abstract The purification of rabbit immunoglobulin molecules expressing kappa (κ) light chains, utilizing the allotypic specificity b4, has been achieved in stages involving isolation of specific antibody, preparation of a solid phase immunoadsorbent of coupled antibody, and subsequent isolation of b4 (κ) IgG. Cellulose trans -2.3-carbonate is shown to be an effective matrix enabling chemical coupling of antibodies and antigens to the support at neutral pH thus preservng immunological activity. The trans -2,3-carbonate derived from microcrystalline cellulose is more effective as a matrix than the trans -2,3-carbonate derived from macroporous cellulose for the chemical coupling of rabbit a1a3/b4 IgG antigen and binding of specific anti-b4 antibody. The microcrystalline celulose carbonate is also more efficient for the coupling of rabbit anti-b4 antibody and the subsequent binding and elution of rabbit b4 (κ) IgG, thus separating immunoglobulin, expressing kappa light chain, from that expressing lambda light chain. The purification technique has potential application in other allotypic systems and antibody- antigen populations.

Phagocyte dysfunction in common variable immune deficiency

ZusammenfassungEs wird über einen 13jährigen Jungen berichtet, der seit der späten Säuglingszeit an häufig wiederkehrenden bakteriellen, enteroviralen und protozoalen Infekten erkrankte. Neben der Erniedrigung der IgG2- IgG3- sowie IgA-Serumspiegel wurden eine Neutropathie und eine partielle Störung der zellulären Infektabwehr gefunden. Ein Folsäuremangel führte zur Panzytopenie und erhöhte damit noch die Infektanfälligkeit. Erst die kombinierte Substitution von Gammaglobulinen und Folsäure vermochte den Circulus vitiosus zu durchbrechen.SummaryThe history of a 13-year old boy is reported who suffered from frequent bacterial, enteroviral, and protozoal infections since late infancy. A decrease in the serum levels of IgG2, IgG3, IgA, a neutrophil dysfunction, and a partial cellular immune deficiency could be demonstrated. A deficiency of folic acid produced a pancytopenia which enhanced the patients susceptibility to infections. The combined substitution of gammaglobulins and folic acid only was able to break this vicious cycle.