Clinical and immunohistochemical data of LV cases
Description
We believe inflammation is a significant factor in LV pathogenesis. Consequently, a preliminary comparative study was conducted on the histopathological slides of certain LV patients admitted to our institution, utilizing immunohistochemistry and other methods to explore this aspect further. We gathered clinical data from 36 patients diagnosed with Livedoid vasculopathy (LV) from the electronic medical records of the Department of Dermatology and Venereology at the First Affiliated Hospital of Guangxi Medical University, spanning January 2019 to December 2023. The dataset included detailed information on patient characteristics such as gender, age, timing of disease onset, predilection sites, disease progression, cutaneous manifestations, preexisting comorbidities, diagnostic laboratory findings, histopathological results, instances of misdiagnosis, and established treatment protocols. All participants had complete and thorough clinical profiles. We make a comprehensive statistical analysis performed on this clinical data. And ten tissue paraffin blocks from randomly selected LV patients were subjected to immunohistochemical analysis to detect the expression of CD3, CD4, CD8, CD20, STAT6, MPO, and MBP. An analysis of immune inflammatory infiltrates was also conducted, using tissue specimens from patients with allergic purpura as a control group. This study confirms that the clinical features of Livedoid vasculopathy (LV) are consistent with previously documented findings. Pathological histological examination identified lymphocytic infiltration as a distinguishing feature of LV. Immunohistochemical analyses revealed that inflammation, particularly involving T-lymphocytes, is central to the pathogenesis of LV. The study also found significant expression of STAT6 in dermal inflammatory cells, suggesting its role in promoting inflammation through the STAT6 signaling pathway.
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1. Case Selection: A total of 10 cases diagnosed with leukocytoclastic vasculitis (LV) by two experienced dermatopathologists from our institution in 2023 were randomly selected, along with 3 cases of hypersensitivity purpura predominantly presenting with leukocytoclastic vasculitis. The corresponding paraffin-embedded tissue blocks were prepared, and multiple sections were cut and baked for subsequent analysis. 2. Laboratory Equipment and Reagents: The experiments were conducted in the dermatopathology immunology laboratory, equipped with all necessary apparatus. All reagents were obtained through legitimate channels and were within their usage period. The reagents included: CD3 antibody (immunohistochemistry), CD4 antibody (immunohistochemistry), CD8 antibody (immunohistochemistry), CD20 antibody (immunohistochemistry), langerin antibody (immunohistochemistry), myeloperoxidase (MPO) antibody (immunohistochemistry), STAT6 antibody (immunohistochemistry), myelin basic protein (MBP) antibody (immunohistochemistry), EDTA antigen retrieval solution, DAB chromogen, and PBS buffer. 3.Procedure: All operations adhered strictly to the protocols outlined in the reagent instructions. The specific experimental steps are as follows: ①Deparaffinization and Rehydration: The baked paraffin sections were placed in fresh xylene, sequentially immersing them in four jars for 5 minutes each at room temperature. After removing excess liquid, the sections were placed in anhydrous ethanol (two jars for 3 minutes each), followed by 95% ethanol (one jar for 3 minutes), and finally 75% ethanol (one jar for 3 minutes). Excess liquid was removed, and the sections were rinsed with distilled water before being placed in freshly prepared PBS buffer. ②Antigen Retrieval: A moderate amount of water was added to a pressure cooker, and a container holding the EDTA antigen retrieval solution (pH 9.0) was placed inside. The mixture was heated on high until boiling. The sections were then immersed in the EDTA solution, covered, and heated under pressure. When the pressure valve began to release steam, the heat was reduced to medium-low and timed for 20 minutes. After this period, the heat source was removed, and the pressure was released before transferring the cooker to cool in cold water. Once the solution reached room temperature, the sections were rinsed in PBS buffer for 2 minutes, repeating this step three times. ③Blocking Endogenous Peroxidase: A 100 µL solution of endogenous peroxidase blocker (3% H2O2) was added, and the sections were incubated at room temperature for 10 minutes. After two washes with distilled water, excess liquid was removed, and a hydrophobic barrier was drawn around the tissue sections using a rabbit anti-viral pen, preventing loss of subsequent reagents. ④Primary Antibody Incubation ⑤Secondary Antibody Incubation ⑥Chromogenic Reaction ⑦Counterstaining ⑧Dehydration, Clearing, and Mounting ⑨Result Interpretation