Clinical and immuno-proteomic approach on Lantana camara pollen allergy-a major health hazard.

BACKGROUND: The incidence of allergic diseases is increasing gradually and is a global burden affecting the socio-economic quality of life. Identification of allergens is the first step towards paving the way for therapeutic interventions against atopic diseases. Our previous investigation figured out that total pollen load correlated significantly with the rise of respiratory allergy in a subtropical city in India. The most dominant pollen responsible for IgE sensitivity in most patients emerged to be from Lantana camara (LC) an obnoxious weed growing in and around suburban areas of West Bengal. In this study, we identified allergenic components from this shrub using an immunoproteomic approach. METHODS: Determination of dominant pollen species was done using aerobiological sampling during two consecutive years and correlated with hospitalization and skin prick test. Serum was collected from LC positive patients and checked for in vitro allergenicity using ELISA and Histamine assay. Total proteome was profiled in SDS-PAGE, 2D PAGE and immunoblotted to detect IgE binding proteins which were further identified using mass spectrometry. RESULTS: Lantana camara pollen emerged as a significant contributor from the correlation study with hospital admission of the respiratory allergy sufferers and its extract demonstrated an elevated IgE response in ELISA and histamine release assay tests. Five IgE reactive bands/zones were observed in 1D blot which resolved to 12 allergo-reactive spots in the 2D blot. Mass spectrometric analysis identified nine spots that grouped into four diverse proteins. Pathogenesis-related Thaumatin-like protein was found to be one of the major allergens in Lantana camara. CONCLUSIONS: This is to our knowledge the first attempt to identify allergens from Lantana camara using a proteomic approach. The allergens identified thereof can be used to prepare hypoallergenic vaccine candidates and design immunotherapy trials against LC pollen and other aeroallergen carriers which are cross-reactive and harbor similar proteins.

Mass spectrometry-based identification of allergens from Curvularia pallescens, a prevalent aerospore in India.

The worldwide prevalence of fungal allergy in recent years has augmented mining allergens from yet unexplored ones. Curvularia pallescens (CP) being a dominant aerospore in India and a major sensitiser on a wide range of allergic population, pose a serious threat to human health. Therefore, we aimed to identify novel allergens from CP in our present study. A cohort of 22 CP-sensitised patients was selected by positive Skin prick grade. Individual sera exhibited elevated specific IgE level and significant histamine release on a challenge with antigenic extract of CP. First gel-based profiling of CP proteome was done by 1- and 2-dimensional gel. Parallel 1- and 2-dimensional immunoblot were performed applying individual as well as pooled patient sera. Identification of the sero-reactive spots from the 2-dimensional gel was found to be challenging as CP was not previously sequenced. Hence, mass spectrometry-based proteomic workflow consisting of conventional database search was not alone sufficient. Therefore, de novo sequencing preceded homology search was implemented for further identification. Altogether 11 allergenic proteins including Brn-1, vacuolar protease, and fructose-bis-phosphate aldolase were identified with high statistical confidence (p<0.05). This is the first study to report on any allergens from CP. This kind of proteome-based analysis provided a catalogue of CP allergens that would lead an improved way of diagnosis and therapy of CP-related allergy.

Biomonitoring of pollen grains of a river bank suburban city, Konnagar, Calcutta, India, and its link and impact on local people.

INTRODUCTION AND OBJECTIVES: Pollen grains released by plants are dispersed into the air and can become trapped in human nasal mucosa, causing immediate release of allergens triggering severe Type 1 hypersensitivity reactions in susceptible allergic patients. Recent epidemiologic data show that 11-12% of people suffer from this type of disorders in India. Hence, it is important to examine whether pollen grains have a role in dissipating respiratory problems, including allergy and astma, in a subtropical suburban city. MATERIALS AND METHODS: Meteorological data were collected for a period of two years, together with aerobiological sampling with a Burkard sampler. A pollen calendar was prepared for the city. A health survey and the hospitalization rate of local people for the above problems were documented following statistical analysis between pollen counts and the data from the two above-mentioned sources. Skin Prick Test and Indirect ELISA were performer for the identification of allergenic pollen grains. RESULTS: Bio-monitoring results showed that a total of 36 species of pollen grains were located in the air of the study area, where their presence is controlled by many important meteorological parameters proved from SPSS statistical analysis and by their blooming periods. Statistical analysis showed that there is a high positive correlation of monthly pollen counts with the data from the survey and hospital. Biochemical tests revealed the allergic nature of pollen grains of many local species found in the sampler. CONCLUSIONS: Bio-monitoring, together with statistical and biochemical results, leave no doubt about the role of pollen as a bio-pollutant. General knowledge about pollen allergy and specific allergenic pollen grains of a particular locality could be a good step towards better health for the cosmopolitan suburban city.