Immunohistochemistry - The Ageless Biotechnology


ND, not done, (+), focally positive.

Reactivities of various pathogens to commercial rabbit antisera against B. cereus, BCG, T. pallidum, and E. coli are summarized.

patients [33]. Figure 11 illustrates the autopsied liver of a Japanese female aged 70's complicated by S. marcescens septicemia. Colonies of Gram-negative microbes were observed in the sinusoid of the liver. Comparative immunostaining disclosed positivity for both E. coli antigens and T. pallidum antigens, in addition to strong reactivity to monoclonal antibody 2D7/1 against E. coli LPS (Abcam) [19].

Serratia marcescens septicemia forming colonies in the sinusoid of autopsied liver (left upper, H&E; right upper, E. coli LPS; left lower, E. coli antigens; right lower, T. pallidum antigens). Colonies of Gram-negative microbes reveal positivity for both E. coli antigens and T. pallidum antigens, in addition to strong reactivity to

Low-Specificity and High-Sensitivity Immunostaining for Demonstrating Pathogens…

Intestinal spirochetosis is caused by localized infection of Brachyspira aalborgi or

immunostaining with T. pallidum monoclonal antibody (J010J), and the spirochete was also proven by immunostaining with antisera against T. pallidum, BCG, E. coli, and B. cereus, as shown in Figure 12 [13, 19]. Antisera against Leptospira and

Non-tuberculous mycobacteria (Mycobacterium avium-intracellulare) in a caseous necrotic lesion of the lung were demonstrated not only by BCG antiserum but also by T. pallidum antiserum, as illustrated in Figure 13 [19]. BCG antiserum was

Gangrenous lesions in the extremity caused by lethal Vibrio vulnificus infection

(flesh-eating disease) microscopically show active growth of Gram-negative

also strongly reactive with amorphous background substances, probably

B. pilosicoli on the colonic mucosa. Basophilic brush border-like structures are recognized in H&E-stained sections [34]. It has been clarified that the brush border-like structures by zoonotic B. pilosicoli are much longer than those by B. aalborgi [35]. The surface-adherent bacteria were clearly visualized by

4.3 Application to intestinal spirochetosis

monoclonal antibody 2D7/1 against E. coli LPS.

DOI: http://dx.doi.org/10.5772/intechopen.85055

Figure 11.

Helicobacter pylori also cross-reacted to the pathogen.

4.4 Cross-reactivity to acid-fast bacilli

representing decayed bacterial proteins.

81

4.5 Application to Vibrio vulnificus infection

#### Table 3.

Reactivities of various microbes to the four kinds of rabbit antibacterial antisera.

#### Figure 10.

Hamster liver in experimental leptospirosis (left, H&E; center, Leptospira antigens; right, E. coli antigens). Liver cell cords are disarranged due to infection of Leptospira interrogans. Not only Leptospira antiserum but also E. coli antiserum decorate spiral-shaped bacteria in the sinusoid. Pathogens phagocytized by activated Kupffer cells are visualized only by Leptospira antiserum.

### 4.2 Application to Serratia septicemia

Serratia marcescens, a red colony-forming Gram-negative rod usually showing low virulence, may cause opportunistic infection in immunosuppressed

Low-Specificity and High-Sensitivity Immunostaining for Demonstrating Pathogens… DOI: http://dx.doi.org/10.5772/intechopen.85055

#### Figure 11.

Serratia marcescens septicemia forming colonies in the sinusoid of autopsied liver (left upper, H&E; right upper, E. coli LPS; left lower, E. coli antigens; right lower, T. pallidum antigens). Colonies of Gram-negative microbes reveal positivity for both E. coli antigens and T. pallidum antigens, in addition to strong reactivity to monoclonal antibody 2D7/1 against E. coli LPS.

patients [33]. Figure 11 illustrates the autopsied liver of a Japanese female aged 70's complicated by S. marcescens septicemia. Colonies of Gram-negative microbes were observed in the sinusoid of the liver. Comparative immunostaining disclosed positivity for both E. coli antigens and T. pallidum antigens, in addition to strong reactivity to monoclonal antibody 2D7/1 against E. coli LPS (Abcam) [19].

## 4.3 Application to intestinal spirochetosis

Intestinal spirochetosis is caused by localized infection of Brachyspira aalborgi or B. pilosicoli on the colonic mucosa. Basophilic brush border-like structures are recognized in H&E-stained sections [34]. It has been clarified that the brush border-like structures by zoonotic B. pilosicoli are much longer than those by B. aalborgi [35]. The surface-adherent bacteria were clearly visualized by immunostaining with T. pallidum monoclonal antibody (J010J), and the spirochete was also proven by immunostaining with antisera against T. pallidum, BCG, E. coli, and B. cereus, as shown in Figure 12 [13, 19]. Antisera against Leptospira and Helicobacter pylori also cross-reacted to the pathogen.
