Intest Res.  2018 Jul;16(3):426-435. 10.5217/ir.2018.16.3.426.

Selective M1 macrophage polarization in granuloma-positive and granuloma-negative Crohn's disease, in comparison to intestinal tuberculosis

Affiliations
  • 1Department of Pathology, All India Institute of Medical Sciences, New Delhi, India.
  • 2Department of Gastroenterology and Human Nutritions, All India Institute of Medical Sciences, New Delhi, India. vineet.aiims@gmail.com dhiraj@icgeb.res.in
  • 3Cellular Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India. vineet.aiims@gmail.com dhiraj@icgeb.res.in

Abstract

BACKGROUND/AIMS
Classical M1 macrophage activation exhibits an inflammatory phenotype while alternative M2 macrophage activation exhibits an anti-inflammatory phenotype. We aimed to determine whether there are discriminant patterns of macrophage polarization in Crohn's disease (CD) and intestinal tuberculosis (iTB).
METHODS
Colonic mucosal biopsies from 29 patients with iTB, 50 with CD, and 19 controls were examined. Dual colored immunohistochemistry was performed for iNOS/CD68 (an M1φ marker) and CD163/CD68 (an M2φ marker), and the ratio of M1φ to M2φ was assessed. To establish the innate nature of macrophage polarization, we analyzed the extent of mitochondrial depolarization, a key marker of inflammatory responses, in monocyte-derived macrophages obtained from CD and iTB patients, following interferon-γ treatment.
RESULTS
M1φ polarization was more prominent in CD biopsies (P=0.002) than in iTB (P=0.2) and control biopsies. In granuloma-positive biopsies, including those in CD, M1φ predominance was significant (P=0.001). In iTB, the densities of M1φ did not differ between granuloma-positive and granuloma-negative biopsies (P=0.1). Interestingly, higher M1φ polarization in CD biopsies correlated with high inflammatory response exhibited by peripheral blood-derived monocytes from these patients.
CONCLUSIONS
Proinflammatory M1φ polarization was more common in colonic mucosa of CD patients, especially in the presence of mucosal granulomas. Further characterization of the innate immune system could help in clarifying the pathology of iTB and CD.

Keyword

Crohn disease; Tuberculosis; Macrophage; M1 macrophage; M2 macrophage

MeSH Terms

Biopsy
Colon
Crohn Disease*
Granuloma
Humans
Immune System
Immunohistochemistry
Macrophage Activation
Macrophages*
Monocytes
Mucous Membrane
Pathology
Phenotype
Tuberculosis*

Figure

  • Fig. 1 (A, B) Colonic biopsies showed more proinflammatory macrophages, than the light brown CD68 positive macrophages in CD (M1 macrophages-reddish brown, black arrows & CD68 positive macrophages, green arrows; A ×40; B ×100). (C) Figure shows confluent tubercular granulomas infiltrated by both M1 macrophages (inset, black arrows) and CD68 positive macrophages (green arrows) (×200). (D, E) Figures show iNOS/CD68 positive M1 macrophages in both superficial and deeper parts of mucosa (black arrows), as well as CD163/CD68 positive M2 macrophages (green arrows) in the deeper part of mucosa in CD (D, ×100; E, ×400). (F) Figure shows polarization of M2 macrophages in mucosal peri-cryptal granuloma of CD (green arrow) (×200).

  • Fig. 2 Peripheral blood mononuclear cells (PBMCs) from 5 CD and 8 intestinal tuberculosis (iTB) patients were differentiated into MDM's using M-CSF (50 ng/mL, 5–7 days). Differentiated macrophages were activated by interferon γ (IFN-γ; 100 U/mL, 12 hours) and stained with JC-1 dye (2 µM) before analyzing the samples in a flow cytometer. The ratio of mean fluorescence intensity in the red to that of green channel is plotted here for each sample. The differences were found significant using Mann-Whitney rank test (P<0.05). MDM, monocyte-derived macrophages; M-CSF, macrophage colony stimulation factors.


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