I need some assistance with these assignment. immunological basis of disease Thank you in advance for the help! a. Comparison of bronchial tissues from asthmatic and non-asthmatic patients by bronchial examination and biopsy. The immunohistochemistry of the proximal airway of bronchial biopsies was conducted by staining them with monoclonal antibodies against smooth muscle actin, and tryptase for mast cells, followed by image analysis.

b. Measuring the effect of the coculturing of mast cells and ASM on the production of smooth muscle actin and TGF-β. Pure cultures of human lung mast cells and airway smooth muscle were produced. ASM cells were labeled with mABs for smooth muscle actin (SMA). ASM cells were cultured with human mast cells, and mast cells that have been sensitized with protease inhibitor and lactoferrin (to inhibit the action of tryptase, a protease produced by mast cells). Under these coculture conditions, the production of smooth muscle actin protein and gene expression were determined periodically for one week using immunofluorescence, flow cytometry, and quantitative real-time PCR. Cell-free supernatants of the cocultures were analyzed for the production of TGF-β1 and TGF-β2 using ELISA. TGF-β ( transforming growth factor-beta) is a protein that controls cell proliferation and differentiation

c. The effect of the addition of recombinant TGF-β on ASM proliferation and contractile activity of ASM pure cultures was also measured by examining SMA protein expression by flow cytometry, and ASM contractile activity by collagen gel analysis.

3. Choose ONE immune-based method used in the paper and, with reference to the paper and the literature, describe this method including its underlying principles. (Use diagrams where appropriate). (No more than 500 words excluding diagram(s)) (30%).

An immune-based method used in the paper was ELISA or enzyme-linked immunosorbent assay. In the paper being reviewed, ELISA was used to quantify the expression of TGF-β1 and TGF-β2 in supernates of ASM and mast cell cultures.

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