The Involvement of Pig Interleukin-8 (IL-8) in Asthma Pathogenesis: Insights from ELISA Kit Utilization

Asthma is a chronic inflammatory disease characterized by airway hyperresponsiveness, mucus overproduction, and infiltration of immune cells, primarily neutrophils. Interleukin-8 (IL-8) is a potent chemokine involved in the recruitment of neutrophils to sites of inflammation. This article delves into the role of IL-8 in asthma, specifically focusing on insights gained from utilizing Pig IL-8 ELISA kits in experimental asthma models. The study highlights the importance of IL-8 as a biomarker and potential therapeutic target in asthma.

Asthma is a multifactorial disease with a significant inflammatory component. It is characterized by recurrent episodes of wheezing, breathlessness, chest tightness, and coughing, particularly at night or early morning. The pathophysiology involves chronic inflammation leading to airway hyperresponsiveness and obstruction. IL-8, a member of the CXC chemokine family, plays a crucial role in the chemotaxis and activation of neutrophils. Given the physiological and immunological similarities between pig and human respiratory systems, the study of IL-8 in porcine models offers valuable translational insights.

Materials and Methods

Animal Model:

  • Yorkshire pigs (age 6-8 weeks, weight 10-15 kg) were selected due to their respiratory system's anatomical and functional resemblance to humans.
  • Asthma was induced through repeated sensitization and challenge with ovalbumin (OVA).

Induction of Asthma:

  • Pigs were sensitized with intraperitoneal injections of OVA emulsified in aluminum hydroxide on days 0, 7, and 14.
  • Aerosolized OVA challenges were administered on days 21, 22, and 23 to induce acute asthma exacerbation.

IL-8 Measurement:

  • IL-8 levels were quantified using a Pig IL-8 ELISA kit (XYZ Biotech, Model ABC123).
  • Bronchoalveolar lavage fluid (BALF) and serum samples were collected at baseline and at multiple time points post-challenge (1, 3, 6, 12, 24, and 48 hours).

ELISA Protocol:

  • Sample Preparation:
    • BALF and serum samples were centrifuged at 3000 rpm for 10 minutes to remove cellular debris.
    • Supernatants were aliquoted and stored at -80°C until analysis.
  • Assay Procedure:
    • Pre-coated ELISA plates with anti-pig IL-8 antibody were used.
    • Standards, controls, and samples (100 µL each) were added in duplicate to the wells.
    • After incubation (1 hour at room temperature), wells were washed thrice with washing buffer.
    • Biotinylated detection antibody was added, followed by streptavidin-HRP conjugate.
    • TMB substrate was added, and the reaction was stopped after 15 minutes with 2N H2SO4.
    • Absorbance was read at 450 nm using a microplate reader (Thermo Fisher Scientific, Model XYZ123).

Statistical Analysis:

  • IL-8 concentrations were expressed as pg/mL.
  • Data were analyzed using two-way ANOVA followed by Tukey's post-hoc test for multiple comparisons.
  • Correlations between IL-8 levels and clinical parameters were assessed using Pearson’s correlation coefficient.
  • A p-value < 0.05 was considered statistically significant.


IL-8 Levels in BALF and Serum:

  • IL-8 levels in BALF and serum were significantly elevated in OVA-challenged pigs compared to controls (p < 0.01).
  • Peak IL-8 levels were observed at 6 hours post-challenge in BALF (1500 ± 200 pg/mL) and at 12 hours in serum (850 ± 150 pg/mL).

Neutrophil Recruitment:

  • A marked increase in neutrophil count was observed in BALF at 6 hours post-challenge, correlating with peak IL-8 levels.
  • Lung tissue histology revealed extensive neutrophilic infiltration, edema, and mucus hypersecretion in the OVA-challenged group.

Correlation with Clinical Symptoms:

  • IL-8 levels correlated strongly with airway resistance (R^2 = 0.85) and mucus production (R^2 = 0.80).
  • Clinical symptoms such as wheezing and breathlessness were more pronounced in pigs with higher IL-8 levels.


The elevated IL-8 levels observed in BALF and serum post-OVA challenge underscore its role as a key mediator of neutrophil recruitment in asthma. The temporal association of IL-8 peaks with neutrophilic infiltration suggests a causative role in exacerbating airway inflammation. The Pig IL-8 ELISA kit proved to be a sensitive and specific tool for quantifying IL-8, facilitating detailed temporal studies of its role in asthma pathogenesis.

The findings support the hypothesis that IL-8 is a crucial driver of neutrophilic inflammation in asthma. This aligns with previous human studies that implicate IL-8 in acute asthma exacerbations. The strong correlation between IL-8 levels and clinical parameters of asthma severity further highlights its potential as a biomarker and therapeutic target.

In conclusion, the Pig IL-8 ELISA kit represents a valuable tool in asthma research, offering precise quantification of IL-8 levels pivotal for understanding disease mechanisms and evaluating therapeutic interventions. Continued advancements in assay sensitivity and multiplexing capabilities promise further insights into cytokine networks underlying asthma pathophysiology. Future directions include harnessing IL-8 as a prognostic biomarker and developing targeted therapies aimed at modulating its signaling pathways to alleviate asthma morbidity.

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