5.2.3.1 Hemorrhagic varicella

Biopsy was taken from the vesicular skin lesion of lethal hemorrhagic varicella (small pox). The patient was a young boy suffering from intractable acute

5.2.4 Protozoan infection

ous leishmaniasis.

Figure 31.

95

5.2.4.2 Acanthamebic keratitis

5.2.4.1 Cutaneous leishmaniasis

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

Figure 31 demonstrates microscopic biopsy features of African-type cutaneous

Another form of cutaneous leishmaniasis, Indian type, is caused by L. tropica [55]. Solitary ulcer formation on the skin is characteristic of this clinical form. An aged Japanese patient stayed in India for months, and skin biopsy was performed in a Japanese hospital. Round pathogens were visualized in the cytoplasm of macrophages in the dermis by the patient's own serum (Figure 32) [22, 23]. Of note is that no cross-reactive staining was noted between the African and Indian types. Namely, the patient serum showed a very high specificity to the respective types of cutane-

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

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

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.

soft contact lens user, complained of eye pain, and under the diagnosis of

leishmaniasis (Leishmania major infection) [54]. Japanese men, a group of 5, volunteered afforestation on the Saharan dessert in the Republic of Mali. Multiple ulcers were formed on the arms of all the members of the group. At the end of the work day, a large swarm of sandflies rushed at them. In a Japanese hospital, biopsy was taken from an ulcer on the arm of one of the members. The dermis was heavily infiltrated by macrophages full of granular microbes. The patient's own serum clearly demonstrated round pathogens in the cytoplasm of macrophages. In the biopsied older lesion seen in his friend, fewer pathogens were identified with

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

immunostaining using the same serum [4, 8, 20–23].

#### Figure 29.

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 reacts to the pathogen.

lymphoblastic leukemia treated with chemotherapy and bone marrow transplantation [52]. The 1:500 diluted serum of another adult patient with a recent 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 substance [53].

#### Figure 30.

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.

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

#### 5.2.4 Protozoan infection

## 5.2.4.1 Cutaneous leishmaniasis

Figure 31 demonstrates microscopic biopsy features of African-type cutaneous leishmaniasis (Leishmania major infection) [54]. Japanese men, a group of 5, volunteered afforestation on the Saharan dessert in the Republic of Mali. Multiple ulcers were formed on the arms of all the members of the group. At the end of the work day, a large swarm of sandflies rushed at them. In a Japanese hospital, biopsy was taken from an ulcer on the arm of one of the members. The dermis was heavily infiltrated by macrophages full of granular microbes. The patient's own serum clearly demonstrated round pathogens in the cytoplasm of macrophages. In the biopsied older lesion seen in his friend, fewer pathogens were identified with immunostaining using the same serum [4, 8, 20–23].

Another form of cutaneous leishmaniasis, Indian type, is caused by L. tropica [55]. Solitary ulcer formation on the skin is characteristic of this clinical form. An aged Japanese patient stayed in India for months, and skin biopsy was performed in a Japanese hospital. Round pathogens were visualized in the cytoplasm of macrophages in the dermis by the patient's own serum (Figure 32) [22, 23]. Of note is that no cross-reactive staining was noted between the African and Indian types. Namely, the patient serum showed a very high specificity to the respective types of cutaneous leishmaniasis.
