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Association of viable Mycobacterium leprae with Type 1 reaction in leprosy
Mrudula Prakash Save
  Mrudula Prakash Save
Affiliation
The Foundation for Medical Research, Worli, Mumbai- 400 018, India
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a
Anju Rajaram Dighe
  Anju Rajaram Dighe
Affiliation
The Foundation for Medical Research, Worli, Mumbai- 400 018, India
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a
Mohan Natrajan
  Mohan Natrajan
Affiliation
National JALMA Institute for Leprosy and other Mycobacterial Diseases (NJIL & OMD). Tajganj, Agra- 282001, India (mohannatrajan13@gmail.com)
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b
Vanaja Prabhakaran Shetty
  Vanaja Prabhakaran Shetty
Affiliation
The Foundation for Medical Research, Worli, Mumbai- 400 018, India
Correspondence to: Vanaja P Shetty, The Foundation for Medical Research, Worli, Mumbai- 400 018, India (Tel: +91-022-24934989; Fax: +91-022-24932876; e-mail: fmr@fmrindia.org)
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a,*

The working hypothesis is that, viable Mycobacterium leprae (M. leprae) play a crucial role in the precipitation of Type 1 reaction (T1R) in leprosy.

Material and Methods:

A total of 165 new multibacillary patients were studied. To demonstrate presence of viable M. leprae in reactional lesion (T1R+), three tests were used concurrently viz. growth in the mouse foot pad (MFP), immunohistochemical detection of M. leprae secretory protein Ag85, and 16s rRNA - using in situ RT- PCR. Mirror biopsies and non reactional lesions served as controls (T1R–).

 Findings:

A significantly higher proportion of lesion biopsy homogenates obtained at onset, from T1R(+) cases have shown unequivocal growth in MFP, proving the presence of viable bacteria, as compared to T1R(–) (P < 0.005). In contrast, few Mirror biopsies were positive in both T1R(+) and T1R(–).

With respect to Ag85, while the overall positivity was higher in T1R(+) (74%), however the intensity of staining (Grade ≥ 2+) was disproportionately higher in T1R(+) BT–BB lesions 11/20 (55%).

In the rebiopsies obtained during a repeat episode of T1R, Ag 85 as well as 16s rRNA, positivity (62% & 100%) was higher in T1R(+).

It is inferred therefore ‘viable’ bacteria are an essential component in T1R and difference in the quality of bacilli, not the quantity or the ratio of dead to viable play a role in the precipitation of T1R.

In conclusion, the findings show that ‘metabolically active’ M. leprae is a component/prerequisite and the secretory protein Ag 85, might be the trigger for precipitation of T1R.
Keywords
Leprosy Viable M. leprae Type 1 reaction Mouse Foot Pad Ag85 16s r-RNA in situ RT-PCR
Cite this article
Mrudula Prakash Save, Anju Rajaram Dighe, Mohan Natrajan, Vanaja Prabhakaran Shetty;
Association of viable Mycobacterium leprae with Type 1 reaction in leprosy; Leprosy Review; 2016; 87; 1; 78-92; DOI: 10.47276/lr.87.1.78
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DOI: 10.47276/lr.87.1.78

articleview.journal_title: Leprosy Review
articleview.publisher: Lepra
articleview.publisher_location: Colchester, UK
articleview.article_date_pre accepted on 03 November 2015 and articleview.pub_on 01 March 2016
© Lepra 2016
LEPROSY
Leprosy Review
0305-7518
British Leprosy Relief Association
Colchester, UK
07-8092
0305-7518/16/064053+15
10.47276/lr.87.1.78
Original Papers
Association of viable Mycobacterium leprae with Type 1 reaction in leprosy
SaveMrudula Prakash
a
DigheAnju Rajaram
a
NatrajanMohan
b
ShettyVanaja Prabhakaran
a
a
The Foundation for Medical Research, Worli, Mumbai- 400 018, India
b
National JALMA Institute for Leprosy and other Mycobacterial Diseases (NJIL & OMD). Tajganj, Agra- 282001, India ()
Correspondence to: Vanaja P Shetty, The Foundation for Medical Research, Worli, Mumbai- 400 018, India (Tel: +91-022-24934989; Fax: +91-022-24932876; e-mail: )
01032016
87
1
78
92
03112015
© Lepra
2016
The working hypothesis is that, viable Mycobacterium leprae (M. leprae) play a crucial role in the precipitation of Type 1 reaction (T1R) in leprosy.
Material and Methods:
A total of 165 new multibacillary patients were studied. To demonstrate presence of viable M. leprae in reactional lesion (T1R+), three tests were used concurrently viz. growth in the mouse foot pad (MFP), immunohistochemical detection of M. leprae secretory protein Ag85, and 16s rRNA - using in situ RT- PCR. Mirror biopsies and non reactional lesions served as controls (T1R–).
Findings:
A significantly higher proportion of lesion biopsy homogenates obtained at onset, from T1R(+) cases have shown unequivocal growth in MFP, proving the presence of viable bacteria, as compared to T1R(–) (P < 0.005). In contrast, few Mirror biopsies were positive in both T1R(+) and T1R(–).
With respect to Ag85, while the overall positivity was higher in T1R(+) (74%), however the intensity of staining (Grade ≥ 2+) was disproportionately higher in T1R(+) BT–BB lesions 11/20 (55%).
In the rebiopsies obtained during a repeat episode of T1R, Ag 85 as well as 16s rRNA, positivity (62% & 100%) was higher in T1R(+).
It is inferred therefore ‘viable’ bacteria are an essential component in T1R and difference in the quality of bacilli, not the quantity or the ratio of dead to viable play a role in the precipitation of T1R.
In conclusion, the findings show that ‘metabolically active’ M. leprae is a component/prerequisite and the secretory protein Ag 85, might be the trigger for precipitation of T1R.
LeprosyViable M. lepraeType 1 reactionMouse Foot PadAg8516s r-RNAin situ RT-PCR