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Toll-Like Receptors 7, 8, and 9 Expression and Function in Primary Human Cervical Cancer Langerhans Cells: Evidence of Anergy
  1. Mahesh M. Kumar, MSc*,
  2. Sreenivas Adurthi, PhD*,
  3. Surya Ramachandran, PhD,
  4. Geetashree Mukherjee, MD,
  5. Omana Joy, PhD§,
  6. H. Krishnamurthy, PhD,
  7. Sudhir Krishna, PhD,
  8. U. D. Bafna, MD,
  9. Devi K. Uma, MD and
  10. R. S. Jayshree, PhD*
  1. *Department of Microbiology, Kidwai Memorial Institute of Oncology, Bangalore;
  2. Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram;
  3. Department of Pathology, Kidwai Memorial Instituteof Oncology, Bangalore;
  4. §FACS Central Facility, Indian Institute of Science, Bangalore, Karnataka; and
  5. National Center for Biological Sciences, Bangalore, Karnataka; and
  6. Department of Gynecology, Kidwai Memorial Institute of Oncology, Bangalore, Karnataka, India.
  1. Address correspondence and reprint requests to R. S. Jayshree, PhD, Department of Microbiology, Room No. 113, 1st Floor, Main Block, Kidwai Memorial Institute of Oncology, Hosur Rd, Bangalore 560029, India. E-mail: microjayshree{at}


Objective Human papillomavirus oncoproteins E6 and E7 down modulate Toll-like receptor (TLR) 9 expression in infected keratinocytes. We explored the status of expression and function of TLR7, TLR8, and TLR9 in primary human Langerhans cells (LCs) isolated from cervical tumors.

Methodology Single-cell suspensions were made from fresh tissues of squamous cell carcinoma (International Federation of Gynecology and Obstetrics stage IB2); myeloid dendritic cells were purified using CD1c magnetic activated cell separation kits. Langerhans cells were further flow sorted into CD1a+CD207+ cells. Acute monocytic leukemia cell line THP-1–derived LCs (moLCs) formed the controls. mRNA from flow-sorted LCs was reverse transcribed to cDNA and TLR7, TLR8, and TLR9 amplified. Monocyte-derived Langerhans cells and cervical tumor LCs were stimulated with TLR7, TLR8, and TLR9 ligands. Culture supernatants were assayed for interleukin (IL) 1β, IL-6, IL-10, IL-12p70, interferon (IFN) α, interferon γ, and tumor necrosis factor (TNF) α by Luminex multiplex bead array. Human papillomavirus was genotyped.

Results We have for the first time demonstrated that the acute monocytic leukemia cell line THP-1 can be differentiated into LCs in vitro. Although these moLCs expressed all the 3 TLRs, tumor LCs expressed TLR7 and TLR8, but uniformly lacked TLR9. Also, moLCs secreted IL-6, IL-1β, and tumor necrosis factor α to TLR8 ligand and interferon α in response to TLR9 ligand; in contrast, tumor LCs did not express any cytokine to any of the 3 TLR ligands. Human papillomavirus type 16 was one of the common human papillomavirus types in all cases.

Conclusions Cervical tumor LCs lacked TLR9 expression and were functionally anergic to all the 3: TLR7, TLR8, and TLR9 ligands, which may play a crucial role in immune tolerance. The exact location of block(s) in TLR7 and TLR8 signaling needs to be investigated, which would have important immunotherapeutic implications.

  • Langerhans cells
  • TLR7
  • TLR8
  • TLR9
  • Cervical cancer
  • CpG ODN
  • Human papillomavirus type 16; Immune tolerance
  • Toll-like receptor

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  • Support was received from Council of Scientific and Industrial Research, New Delhi (Senior Research Fellowship to M.M.K.) and the Indian Council of Medical Research, New Delhi (Senior Research Fellowship to S.A.). Financial assistance was also received from the Department of Biotechnology, New Delhi, for the project entitled, “TLR Expression Profile in Langerhans and Dendritic Cells in Invasive Cervical Cancer, Cervical Intraepithelial Neoplasia and Chronic Cervicitis” (no. BT/PR5353A/MED/14/621/2004).

  • The authors declare no conflicts of interest.