Duaine R. Jackola

Evidence of an affinity threshold for IgE‐allergen binding in the percutaneous skin test reaction

Background Atopy is an aberrant immune response involving allergen‐specific IgE production, though serum IgE concentration is not an entirely reliable diagnostic tool, particularly for epidemiological and genetic studies. There is no clear correlation between IgE and other indicators of atopy such as skin prick tests (SPT)s, and physiological associations are difficult to justify in cases with detectable IgE but negative SPT results.

Altered allergen binding capacities of Amb a 1-specific IgE and IgG4 from ragweed-sensitive patients receiving immunotherapy

BACKGROUND The mechanisms for the effectiveness of allergen immunotherapy (IT) are not well understood. The binding potential for immunoglobulins is a function of both antibody concentration and affinity (K(A)). PURPOSE The purpose was to perform a cross-sectional preliminary study to investigate any differences in allergen-specific antibody affinity and concentration following ragweed immunotherapy by introducing a new concept of antibody binding capacity ([Ig] X K(A)). METHODS The binding capacity of allergen-specific IgE and IgG4 was determined for ragweed-allergic individuals undergoing ragweed immunotherapy and compared with the capacity of ragweed-specific IgE and IgG4 for allergic individuals not receiving immunotherapy. RESULTS The mean binding capacity for IgG4 after long-term immunotherapy was 1.6 log units higher (P < .0001) than for individuals not receiving IT. The binding capacity for allergen-specific IgE was 1.2 log units lower following long-term immunotherapy (P < .0001) compared with individuals not receiving ragweed IT. CONCLUSIONS We hypothesize that a primary effect of immunotherapy is to increase IgG4 binding capacity and concomitantly decrease IgE binding capacity.

CD14 Promoter Polymorphisms in Atopic Families: Implications for Modulated Allergen-Specific Immunoglobulin E and G1 Responses

Background: CD14 promoter DNA sequence polymorphisms for the endotoxin receptor gene have been implicated in modulating allergen-specific immunoglobulin (Ig)E responses in randomly selected individuals with atopy. We sought to determine if a single nucleotide polymorphism in the CD14 promoter region is associated with atopy in atopic families, and to assess its influence on serum levels of CD14 and allergen-specific IgE and IgG1 responses. Methods: We screened 367 members of 91 Caucasian nuclear families with a history of asthma for pulmonary function by spirometry, including methacholine challenge to detect bronchial hyperreactivity, and atopy by serum total IgE and skin prick test to 14 allergens. The CD14 promoter single nucleotide polymorphism was analyzed in DNA isolated from peripheral blood mononuclear cells to identify C/C, C/T and T/T genotypes. Serum tests were done for soluble CD14 (sCD14) and dust mite-specific antibody (Der p 1-IgG1). Results: Serum sCD14 levels were not associated with clinical phenotypes (asthma, bronchial hyperreactivity or atopy). However, sCD14 levels were inversely related to both allergen-specific IgE and Der p 1-IgG1 production, but only among those with evidence of atopic sensitization. Linear regression analysis, accounting for random family effects, demonstrated a higher production of allergen-specific IgE or Der p 1-IgG1 associated with the T/T genotype and a lower level of specific IgE and IgG1 production associated with sCD14 levels. Conclusions: An element of the innate immune system (CD14) has profound effects upon modulating the acquired allergen-specific immunoglobulin responses among those with an inherited atopic predisposition.

Allergen Skin Test Reaction Patterns in Children (≤10 Years Old) from Atopic Families Suggest Age-Dependent Changes in Allergen-IgE Binding in Early Life

Background: Genetic studies of atopy rely upon evidence of abnormal IgE production, usually elevated total IgE or skin prick test (SPT) reactions. However, these measures may change with subject age. Methods: We screened 1,099 members of atopic families (aged 6–87 years) by serum total IgE and SPT for 14 allergens. For those SPT negative, we screened for Amb a 1- and Der p 1-specific IgE. Der p 1 IgE-Der p 1 allergen binding affinities were done on randomly selected subjects. Results: There were significantly fewer atopics ≤10 years old (69.1%) compared to those >10 years old (75.8%) based upon any SPT-positive result. Children ≤10 years had fewer SPT-positive reactions and smaller SPT wheal reaction areas. Yet, mean total IgE values were comparable to those of the older group. Screens for specific IgE showed no differences in proportions of atopics (≤10 years old = 83.1% and >10 years old = 82.3%). Among those SPT-positive for house dust mite extract, there was a positive correlation between Der p 1 binding affinity and the wheal area of the house dust mite extract. There was a positive correlation between the number of SPT-positive reactions and total IgE for both age groups. However, there was only a significant relationship between SPT-positive wheal area and total IgE for those >10 years old and no apparent relationship between wheal area and total IgE for those ≤10 years old. Conclusion: These results suggest that atopy-specific physiological mechanisms, primarily those involving allergen-IgE binding, change during the earliest years of life.

Random outcomes of allergen‐specific responses in atopic families

Background Allergens are common non‐infectious antigens to which people will mount T cell dependent humoral responses. Among genetically susceptible individuals, an antigen‐specific response results involving the production of allergen‐specific IgE (atopy).

Evidence for expansion of a population of lymphocytes with reduced or absent T3 expression in aged human donors

In a previous report, we described an unusual pattern of T cell associated surface marker expression in unfractionated mononuclear cells from aged donors; an excess of T4 and T8 positive cells relative to T3 positive cells. This study further characterizes these cells on the basis of density, adherence to nylon wool and quantitative expression of cell surface markers. We find that the population of lymphocytes responsible for the unusual surface marker expression is of low density, adheres to nylon wool, and is present in small numbers in young donors. The adherent cells have a reduced quantitative expression of the T3 antigen, no change in the antigen density of T4 and T8, and have increased expression of the T10 antigen. These cells do not have the characteristics of large granular lymphocytes, monocytes, B cells with unusual marker expression, or thymocytes poised for export to peripheral blood. We suggest that these cells, found in increased numbers in aged humans, may represent an expansion of a population of T lymphocytes with absent or reduced T3 antigen expression found normally in smaller numbers in young adults. T lymphocyte antigen receptor density has been quantitatively linked to expression of the T3 antigen. Thus, our results imply that aging may lead to decreased T cell surface antigen density, which may account in part for decline in T cell function with age.

Expression of IgH chain genes in antibody response to ragweed and purified Amb a 1: evidence for differential regulation of isotype production

A survey of the work with Ig response to allergens carried out previously reveals an allergen-specific response both by IgE and all of IgG subclasses. Response of non-sensitive people is characterized by the appearance of a variety of the IgG subclasses. We have reexamined ragweed and Amb a 1 specific Ig response in 54 nonsensitive and 147 atopic or atopic-allergic people using a new inverse sandwich immunoassay allowing discrimination based on antibody affinity. We show that non-sensitive people present no, 0 out of 54, Ig response with affinities higher than Ka 10(7) M(-1). The subpopulation of 66 atopics who never have experienced desensitization responds vigorously and solely (56 out of 66) with genes of the sequence gamma2-alpha2. Only ten showed an additional weak response from gamma1-alpha1. This suggests a possible association between the atopic state and selective activation of part of the gene sequence.

Evidence for lymphocyte chemotaxis toward monocytes during PHA-induced aggregation in vitro

An important, early phenomenon during the development of immune cell interactions in vitro is the formation of multicellular aggregates. We have developed a quantitative assay to determine the kinetics of multicellular aggregate formation within a heterotypic population of cells on a flat surface. This assay follows the time rate of change in the value of an aggregation index for cells in undisturbed culture. For an initial, well-separated population of cells, the index is a minimum and remains at this value if the cells do not move and interact. By contrast, for conditions that promote active cell movement followed by interaction, the index value increases with time. The index, which reflects cells’ relative spatial distributions, is an “indirect enumeration” of the number of cells within aggregates as a function of time.We used this index to follow the aggregative behavior of a population of freshly isolated human peripheral lymphocytes and monocytes. Previous studies have shown that monocytes are centrally located within aggregates and that lymphocytes move to surround monocytes. In order to test if lymphocyte movements are random or directed prior to interactions with monocytes, we formulated a simple model to describe changes in the expected number of cells in an “idealized aggregate” as a function of time. A comparison of the model curves with curves generated from the changes in the aggregation index shows that the best fit derives from a model that involves directed movement of lymphocytes toward monocytes. These results suggest that monocytes produce a chemoattracting agent for lymphocytes for these experimental conditions.