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Antigen protein and nucleic acid coding for said protein

Inactive Publication Date: 2005-12-15
TAKARA HOLDINGS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008] The present inventors have investigated about a method for conferring Candida albicans-infection resistance to mammals, such as mice and rats, which are sensitive to Candida albicans infection. As a result, the present inventors have clarified that such mammals can acquire strong resistance to infection by immunizing the mammals with Candida albicans cells as the antigen in mixture with an adjuvant of incomplete Freund's adjuvant. The present inventors have also clarified that CD4-positive T-cells play a ke

Problems solved by technology

Even in normal individuals, however, this fungus can cause local infectious disease; in females, particularly in pregnant women, it can cause vaginal candidiasis.

Method used

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  • Antigen protein and nucleic acid coding for said protein

Examples

Experimental program
Comparison scheme
Effect test

example 1

Induction of Infection Resistance and Involvement of CD4-Positive T-Cells in Infection Resistance

1) Induction of Infection Resistance

[0108]Candida albicans (C. albicans) TIMM 1768 was cultured overnight with shaking in Sabouraud dextrose medium. Thereafter, cultured cells were harvested by centrifugation and washed with physiological saline. The obtained cells were suspended in physiological saline so as to have concentrations of 1×106, 1×107, and 1×108 cells / ml. An equal volume of IFA (incomplete Freund's adjuvant) was added to each suspension and mixed. This mixture was subcutaneously administered to C57BL / 6 mice at 0.1 ml per animal, thereby immunizing the mice with live Candida cells. After 1 week, the mice were further immunized by subcutaneously administering the same number of live Candida cells prepared in the same manner as above. Therefore, each mouse was twice immunized with 5×104, 5×105, or 5×106 live Candida cells. As a control, a mixture of physiological saline and ...

example 2

Collection of Serum from Mammal Having Infection Resistance, Characteristics Thereof, and Preparation of Various Absorption Sera

1) Collection of Serum from Infection-Resistant Mammal

[0114] Serum was collected from BALB / c mice that were immunized 3 times with 5×106 live C. albicans TIMM 1768 cells in a mixture with CFA (complete Freund's adjuvant) in the same manner as in item 2) of Example 1 by a conventional method at 7 days after final immunization. This resulting serum was used as an anti-Candida serum.

2) Characteristics of Serum from Infection-Resistant Mammal

[0115] The characteristics of the anti-Candida serum obtained in item 1) of Example 2 were studied.

[0116] First, as antigenic components, C. albicans cell wall fraction (CW), cytoplasm fraction (HSS), and cell membrane fraction (LSP) were prepared.

[0117] A loopful of C. albicans TIMM 1768 cells in Sabouraud agar slant culture was transferred to a test tube containing 5 ml of YPD medium (1% by weight yeast extract, 2...

example 3

Screening of C. albicans cDNA Expression Library for Antigenic Proteins

1) Preparation of C. albicans cDNA Expression Library

[0123] Total RNA was extracted and purified from cells to prepare a cDNA expression library for C. albicans TIMM 1768 strain. Specifically, the above strain was cultured at 35° C. in 200 ml of YPD medium, and thereafter the cells were harvested by centrifugation (2,000 rpm, 5 minutes) and washed once with distilled water. The cells were quickly frozen with liquid nitrogen, and the frozen cells were disrupted with a mortar into a powdery form. Total RNA was recovered and purified from this cell powder using an RNA extraction kit (manufactured by Pharmacia). poly(A)+ RNA was prepared using Oligotex™-dT30 (manufactured by Takara Shuzo Co., Ltd.) from this total RNA. cDNA was synthesized from 5 μg of the poly(A)+ RNA using cDNA synthesis kit (manufactured by Takara Shuzo Co., Ltd.). A cDNA library was constructed by ligating the synthesized cDNA to lambda phage...

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Abstract

To provide a novel antigenic protein and a nucleic acid encoding the antigenic protein, which are useful for prophylaxis, treatment and diagnosis of diseases caused by fungi including Candida albicans. An antigenic protein characterized in that the antigenic protein is recognized by antiserum derived from a mammal having Candida albicans-infection resistance; and a nucleic acid encoding an antigenic protein which is recognized by antiserum derived from a mammal having Candida albicans-infection resistance.

Description

[0001] This application is a Divisional of co-pending application Ser. No. 09 / 509,744 filed on Mar. 31, 2000 and for which priority is claimed under 35 U.S.C. § 120. Application Ser. No. 09 / 509,744 is the national phase of PCT International Application No. PCT / JP98 / 04326 filed on Sep. 28, 1998 under 35 U.S.C. § 371. The entire contents of each of the above-identified applications are hereby incorporated by reference. This application also claims priority of Application No. 9-269087 filed in Japan on Oct. 1, 1977 under 35 U.S.C. § 119.TECHNICAL FIELD [0002] The present invention relates to Candida albicans antigenic protein, and a nucleic acid encoding thereof, which are useful for prophylaxis, treatment and diagnosis of fungal infectious disease or allergosis. The present invention further relates to a vector comprising the nucleic acid, and a transformant resulting from transformation with the vector. The present invention further relates to a method for producing the antigenic pro...

Claims

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Application Information

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IPC IPC(8): G01N33/50A61K38/00A61K39/00A61K49/00A61P13/02A61P15/00A61P31/04C07K14/40C07K16/14C12N5/10C12N15/09C12N15/31C12Q1/68G01N33/53G01N33/569
CPCA61K38/00Y10S530/824C07K14/40A61K39/00A61P13/02A61P15/00A61P31/04C12N15/11C12N15/63
Inventor TAKESAKO, KAZUTOHMIZUTANI, SHIGETOSHIENDO, MASAHIROOGAWA, JUNKOOKADO, TAKASHIKATO, IKUNOSHIN
Owner TAKARA HOLDINGS
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