Sarcoidosis (SA) is a multisystem granulomatous disorder of unknown etiology. Infectious organisms, e.g. Mycobacterium tuberculosis, genetic factors and autoimmunity are considered as etiologic agents. Pathologic similarities between SA and tuberculosis (TB) suggest M. tuberculosis heat shock proteins (Mtb-hsp) as causative factors. To test the “mycobacterial” origin of SA, we evaluated the presence of serum anti-Mtb-hsp70, -Mtb-hsp65 and -Mtb-hsp16 antibodies in SA and TB. Analysis of anti-Mtb-hsp antibodies was carried out in 37 patients with SA, 29 patients with TB and 18 healthy individuals by ELISA. Our results show a significantly higher occurrence of anti-Mtb-hsp70 antibodies in SA and TB patients than in controls. The anti-Mtb-hsp65 and -Mtb-hsp16 antibodies occured significantly more often in TB than in controls and SA. We found significantly higher percentages of anti-Mtb-hsp70 and -Mtb-hsp65 antibodies in Stage II compared to Stage I of SA. Analysis of anti-Mtb-hsp antibodies levels revealed significantly higher anti-Mtb-hsp70 antibodies level in SA and TB than in controls. Significantly higher frequency and levels of anti-Mtb-hsp70 than anti-Mtb-hsp65 and -Mtb-hsp16 antibodies was found in SA only. In summary, the frequency and level of anti-Mtb-hsp70 antibodies were comparable between SA and TB but were significantly higher compared to controls and other tested Mtb-hsp. These data may suggest a role for Mtb-hsp70 protein in the pathogenesis of sarcoidosis.