5.2.4.2 Acanthamebic keratitis

lymphoblastic leukemia treated with chemotherapy and bone marrow

history of varicella clearly reacted to the plasma membrane of acantholytic keratinocytes with intranuclear viral inclusion bodies, and the localization pattern was comparable with that of GP-1 antigen revealed by immunostaining using monoclonal antibody C90.2.8 (Figure 30) [21, 23]. The viral intranuclear inclusions remained unreactive. It is evident that antibodies raised in the infected patient were

specific to GP-1 antigen of varicella-zoster virus, an immunodominant viral

Hemorrhagic varicella (left, H&E; center, reactivity with the serum of another patient; right, reactivity with monoclonal antibody C90.2.8 to GP-1 antigen of varicella-zoster virus). Biopsy was taken from a hemorrhagic vesicle, as lethal manifestation of opportunistic varicella-zoster virus infection in a leukemic boy after bone marrow transplantation. The diluted serum of another adult patient with a recent history of varicella clearly reacts to the plasma membrane of acantholytic keratinocytes with unreactive intranuclear viral inclusion

bodies. The localization pattern is comparable with that of GP-1 antigen.

substance [53].

Figure 30.

94

Figure 29.

reacts to the pathogen.

Immunohistochemistry - The Ageless Biotechnology

transplantation [52]. The 1:500 diluted serum of another adult patient with a recent

Cutaneous cryptococcosis (left, H&E; right, reactivity with patient's own serum). Opportunistic skin infection of Cryptococcus neoformans in a treated leukemia case of the young microscopically reveals transparent yeasts floating in mucoid material. Inflammatory reaction is poor. The patient's own serum diluted at 1:10 weakly

> Acanthamebic keratitis is sight-threatening infection of the cornea by the genus Acanthamoeba, mostly seen in contact lens wearers [56]. A young male aged 10's, a soft contact lens user, complained of eye pain, and under the diagnosis of acanthamebic keratitis, superficial keratectomy was done. Acanthamoeba spp. was cultured. As shown in Figure 33, acanthamebic cysts and some trophozoites were identified in the corneal stroma by immunostaining using both the diluted serum of a patient of acanthamebic meningoencephalitis (see Figure 49) and a monoclonal

#### Figure 31.

African-type cutaneous leishmaniasis (left, gross appearance of the forearm; upper panels, the patient of gross photograph; lower panels, an older lesion of his friend; center, H&E; right, reactivity with patient's serum). The Japanese men volunteered afforestation on the Saharan dessert in the Republic of Mali. Biopsy was taken from ulcerated skin lesions. The patient's own serum demonstrates round pathogens (Leishmania major) in the cytoplasm of macrophages infiltrating in the dermis (upper panels). In the older lesion seen in his friend (lower panels), fewer pathogens are observed with immunostaining employing the same serum.

#### Figure 32.

Indian-type cutaneous leishmaniasis (left, gross appearance of the neck; center, H&E; right, reactivity with patient's own serum; inset, Giemsa stain on a stamp cytology preparation). Solitary skin ulcer is characteristic of this clinical form endemic in India. Biopsy specimen from a Japanese patient shows numerous round pathogens (Leishmania tropica) in the cytoplasm of macrophages in the dermis. The diluted patient's own serum visualizes the pathogen. Giemsa preparation demonstrates a pair of the nucleus and kinetoplast (smaller dot) in the protozoan cell.

5.2.4.4 Toxoplasmosis

Figure 34.

Figure 35.

97

of Toxoplasma meningitis (Figure 35) [21, 22].

HIV-negative female with progressive supranuclear palsy.

Cryptogenic and asymptomatic infection of Toxoplasma gondii is common throughout the world. In patients with compromised immune system, particularly in AIDS, the parasite is activated, resulting in lethal Toxoplasma encephalitis. When primary infection occurs in pregnant women without serum antibody, the placental tissue is infected, causing congenital toxoplasmosis in the fetus [59]. Ruptured protozoan cysts of AIDS-associated cerebral toxoplasmosis were reactive with the serum from a healthy individual serologically with high-titer IgG. The same serum decorated trophozoites of T. gondii dispersed in the meningeal space in an aged HIV-negative female with progressive supranuclear palsy, confirming the diagnosis

Toxoplasmosis (upper panels, toxoplasma encephalitis in AIDS; lower panels, toxoplasma meningitis in a non-AIDS patient; left, H&E; right, reactivity with the serum from a healthy individual serologically with high-titer IgG). Ruptured protozoan cysts in AIDS-associated cerebral toxoplasmosis are reactive with the serum. The same serum decorates trophozoites of Toxoplasma gondii (arrows) dispersed in the meningeal space in an aged

Amebic dysentery (left, H&E; center, reactivity with patient's own serum; right, reactivity with monoclonal antibody EHK153 to E. histolytica). Surgically resected colon specimen from an HIV-negative Japanese male demonstrates amebic trophozoites invading the ulcerated colon wall. The patient's own serum visualizes the pathogen, and the localization was comparable with immunostaining using the monoclonal antibody.

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

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

#### Figure 33.

Acanthamebic keratitis (left, H&E; center, reactivity with the serum of patient of acanthamebic meningoencephalitis; right, reactivity with monoclonal antibody ACA5 to Acanthamoeba sp.). Superficial keratectomy specimen from a young contact lens wearer reveals scattered amebic cysts (arrows) in the corneal stroma, and the microbes (cysts) are visualized by immunostaining using both the diluted patient's serum and the monoclonal antibody.

antibody ACA5 to Acanthamoeba spp. (a gift from Prof. Hiroshi Tachibana, Department of Infectious Diseases, Tokai University School of Medicine, Isehara) [57].

#### 5.2.4.3 Amebic dysentery

Amebic dysentery, colonic infection of Entamoeba histolytica, is endemic in areas with poor sanitation and is often associated with AIDS [58]. Surgically resected colon specimen from an HIV-negative Japanese male patient demonstrated amebic trophozoites invading the ulcerated colon wall. As shown in Figure 34, the patient's own serum visualized the pathogen, and the localization was comparable with immunostaining using a monoclonal antibody EHK153 to E. histolytica (a gift from Prof. Yuji Tachibana, Department of Infectious Diseases, Tokai University School of Medicine, Isehara) [4, 20–23].

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

#### Figure 34.

Amebic dysentery (left, H&E; center, reactivity with patient's own serum; right, reactivity with monoclonal antibody EHK153 to E. histolytica). Surgically resected colon specimen from an HIV-negative Japanese male demonstrates amebic trophozoites invading the ulcerated colon wall. The patient's own serum visualizes the pathogen, and the localization was comparable with immunostaining using the monoclonal antibody.
