Bioengineering a humanized acne microenvironment model: Proteomics analysis of host responses to Propionibacterium acnes infection in vivo

Abstract

Acne is a human disease of the sebaceous hair follicle. Unlike humans, most animals produce little or no triglycerides in hair follicles to harbor Propionibacterium acnes a fact that has encumbered the development of novel treatments for acne lesions. Although genetic mutant mice with acne-like skins have been used for screening anti-acne drugs, the mice generally have deficits in immune system that turns out to be inappropriate to generate antibodies for developing acne vaccines. Here, we employed a bioengineering approach using a tissue chamber integrated with a dermis-based celltrapped system (DBCTS) to mimic the in vivo microenvironment of acne lesions. Human sebocyte cell lines were grown in DBCTS as a scaffold and inserted into a perforated tissue chamber. After implantation of a tissue chamber bearing human sebocytes into ICR mice, P. acnes or PBS was injected into a tissue chamber to induce host immune response. Infiltrated cells such as neutrophils and macrophages were detectable in tissue chamber fluids. In addition, a proinflammatory cytokine macrophage-inflammatory protein-2 (MIP-2) was elevated after P. acnes injection. In tissue chamber fluids, 13 proteins including secreted proteins and cell matrix derived from mouse, human cells or P. acnes were identified by proteomics using isotope-coded protein label (ICPL) coupled to nano-LC-MS analysis. After P. acnes infection, four proteins including fibrinogen, α polypeptide, fibrinogen β chain, S100A9, and serine protease inhibitor A3K showed altered concentrations in the mimicked acne microenvironment. The bioengineered acne model thus provides an in vivo microenvironment to study the interaction of host with P. acnes and offers a unique set-up for screening novel anti-acne drugs and vaccines.

Sebum Free Fatty Acids Enhance the Innate Immune Defense of Human Sebocytes by Upregulating β-Defensin-2 Expression

“…We have developed a tissue chamber model integrated with a dermis-based; cell-trapped system to mimic the in vivo microenvironment of acne lesions. A tissue chamber bearing human sebocytes is implanted into ICR mice, and then P. acnes can be injected into the tissue chamber to induce a host immune response (Nakatsuji et al ., 2008). Thus, we anticipate that the tissue chamber model will be a useful animal model for additional studies of the physiological functions of FFAs in the innate immune defense of human sebocytes.…”

Antimicrobial Property of Lauric Acid Against Propionibacterium Acnes: Its Therapeutic Potential for Inflammatory Acne Vulgaris

“…P. acnes is resistant to phagocytes and is able to survive in macrophages (Webster et al , 1985). In our previous data obtained using a tissue chamber model integrated with a dermis-based cell-trapped system to mimic the in vivo microenvironment of acne lesions, injection of living P. acnes into the intradermally implanted tissue chamber attracted Gr-1 + neutrophils and CD11b + macrophages into the chamber (Nakatsuji et al , 2008d) and increased the level of proinflammatory cytokine and macrophage inflammatory protein-2 in the chamber fluid (Nakatsuji et al , 2008a, b). We have recently developed effective vaccines for P. acnes associated inflammation as an alternative treatment for acne.…”

Histone H4 Is a Major Component of the Antimicrobial Action of Human Sebocytes

“…Tryptic peptide digests were analyzed by Nano liquid chromatography-mass spectrometry as described earlier (Nakatsuji et al , 2008). The automated Nano liquid chromatography-mass spectrometry setup consisted of an Agilent 1200 Nano 2D LC system, a switch valve, a C18 trap column (Agilent, Santa Clara, CA), and a capillary reverse-phased column (10 cm in length, 75 µm i.d.)…”