HKMT

Heat-killed Mycobacterium tuberculosis

HKMT is a heat-killed preparation of the avirulent strain of Mycobacterium tuberculosis H37 Ra.

HKMT is sensed by Mincle which recognizes the mycobacterial cell wall glycolipid TDM. HKMT also possesses a large repertoire of TLR2 ligands, such as lipoproteins and lipomannan. Upon HKMT sensing, both Mincle and TLR2 lead to the activation of NF-κB.

References:

1. Ishikawa E. et al., 2009. Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C-type lectin Mincle. 206(13):2879-88.
2. Bhatt K & Salgame P., 2007. Host innate immune response to Mycobacterium tuberculosis. J Clin Immunol 27(4): 347–362.

Specifications

Specificity: Mincle agonist, TLR2 agonist

Working concentration:

• 10 - 100 µg/ml when used as a Mincle ligand
• 100 ng - 10 µg/ml when used as a TLR2 ligand

Contents

Heat-killed preparation of Mycobacterium tuberculosis H37Ra (HKMT) is provided lyophilized and is available in two quantities:

tlrl-hkmt-1:

• 10 mg lyophilized HKMT
• 1.5 ml endotoxin-free water

tlrl-hkmt-5:

• 5 x 10 mg lyophilized HKMT
• 10 ml endotoxin-free water

HKMT is shipped at room temperature

Stored at 4˚C.

Description

HKMT is a heat-killed preparation of the avirulent strain Mycobacterium tuberculosis H37Ra, which was derived from the virulent strain H37. The attenuated M. tuberculosis strain H37Ra is one of the most commonly used controls for M. tuberculosis.

HKMT is sensed by the C-Type lectin Mincle (macrophage-inducible C-type lectin), which recognizes the mycobacterial cell wall glycolipid TDM [1]. HKMT also possesses a large repertoire of TLR2 ligands, such as lipoproteins and lipomannan [2, 3].

Upon HKMT sensing, both Mincle and TLR2 lead to the activation of NF-κB. Specifically, Mincle interacts with the Fc receptor common γ-chain (FcRγ), which triggers intracellular signaling through Syk leading to CARD9-dependent NF-κB activation [4, 5]. HKMT does not activate TLR4 even at high concentrations (up-to 100 µg/ml).

1. Ishikawa, e. et al., 2009. Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C-type lectin Mincle. J. Exp. Med. 206, 2879–2888.
2. Bhatt K & Salgame P., 2007. Host innate immune response to Mycobacterium tuberculosis. J Clin Immunol 27(4): 347–362.
3. Underhill dm. et al., 1999. Toll-like receptor-2 mediates mycobacteria-induced proinflammatory signaling in macrophages. PNAS. 96(25):14459-63.
4. Schoenen H. et al., 2010. Cutting edge: Mincle is essential for recognition and adjuvanticity of the mycobacterial cord factor and its synthetic analog trehalosedibehenate. J Immunol. 184(6):2756-60.
5. Kerscher B. et al., 2013. The Dectin-2 family of C-type lectin-like receptors: an update. Int Immunol. 25(5):271-7.

Details

Responses to M. tuberculosis-derived ligands.
RAW-Blue™ cells, and HEK-Blue™ hMincle, HEK-Blue™ TLR2 or HEK-Blue™ TLR4 cells, which express the human mincle, TLR2 or TLR4 gene, respectively, were stimulated with 10 µg/ml of M. tuberculosis-derived ligands. After 24h incubation, NF-κB activation was assessed by measuring the levels of SEAP using the QUANTI-Blue™ assay.