Infectious Disease Group

[Infectious Diseases]
All infectious diseases such as common cold, bronchitis, pneumonia, cystitis, meningitis, and various purulent wounds are caused by “germs”. “Germs” can be divided into many species such as virus, bacteria, fungi, and parasites and usually cannot be seen by naked eyes. AIDS is the one of the most famous infectious diseases and caused by so called HIV virus. Common cold and pneumonia is mostly caused by viral and bacteria, respectively. Since pathogenic organisms are diverse, all infectious diseases are diagnosed by using various techniques and detection systems and the treatment is also different for each causative microorganisms. In our section, the medical practices, the researches and the educations are being well organized and performed in order to make precious and rapid diagnosis of infectious diseases and treatment.
Since infectious diseases are caused by many kinds of “germs”, it is important to establish the proper diagnosis rapidly, and the prompt diagnosis allows us to select the most effective antibiotics. For example, the microscopic examination is an useful and quick test in order to make a rapid diagnosis, we examine a specimen immediately after its collection from the infected site. This examination allows us to make a rough diagnosis within half an hour, whereas the conventional culture of organisms takes at least three days to detect the certain organisms. We also have a genetic tool for rapid diagnosis of tuberculosis. It takes only a couple of days to make a diagnosis of tuberculosis, which took several months formerly.
Not only developing diagnosis tools or methods, we also take an initiative in the use of newly developed drugs and evaluating the effectiveness of them in clinical trials. Since the clinical trials of investigational drugs are considered as a special task that university hospitals are given to do, we are performing many clinical trials. Needless to say, we are paying great regards to the patient’s benefit so that we never perform a single clinical trial without proper informed consents.
It has been about eighty years since we have been conducting wide varieties of the researches and clinical practices, we believe that we are taking the lead in infectious diseases field in Japan.

[Pulmonary Tuberculosis]

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. The route of infection has been considered airborn; human tubercle bacilli are transmitted by inhalation of droplets of sputum containing the causative organism. Therefore, patients who are diagnosed smear positive of tubercle bacilli should be legally isolated to prevent spreading. In Japan, about 50,000 new cases of tuberculosis are reported every year recently. It is recognized as important problem that patients with acquired immunodeficiency syndrome (AIDS) are particularly susceptible to tuberculosis in the whole world. Today, multi-drug resistant M. tuberculosis is also globally important. In order to prevent relapses of tuberculosis and emergence of drug resistant organisms, if tuberculosis is diagnosed, the patient must receive an appropriate drug therapy. Chemotherapeutic regimens consisting of 3 or 4 anti-tuberculosis drugs are widely accepted, and a long-term therapy lasting for 6 to 9 months is necessary. If the infection of M. tuberculosis is suspected, the patient should receive an examination and proper treatment, following instructions given by a physician.

[Fungal Diseases]

Fungal diseases can be divided into two types such as superficial type and deep-seated type. The lesions of the former type are limited to the skin, whereas those of the latter type involve interior organs. Candidiasis, aspergillosis, and criptococcosis are clinically important deep mycosis. Aspergillosis is predominant recently in autopsy cases in place of candidiasis. Candida albicans normally resides in the oral cavity, gastrointestinal tract, and skin of the healthy humans. However, it causes candidemia when the host defence system is impaired. Aspergillus species and Cryptococcus neoformans are natural inhabitants and they can also cause fatal diseases in immuno-compromised hosts. Particularly, C. neoformans, which can be isolated from feces of pigeons, is transmitted to even healthy humans to cause infection. In recent years, the number of the deep-seated fungal infection cases occurring as opportunistic infections have been increasing in association with the expansion of AIDS, usage of anti-cancer drugs and the organ transplantation cases. Although the organ transplantation is still minor in Japan, it will be performed more and more commonly in the near future. It is obvious that deep-seated fungal infections in organ transplantation patients will be a major clinical concern.

Researchs

[Animal Models for Infectious Disease Research]
We have established various animal models that mimic infectious diseases in humans such as chronic respiratory tract infection with Pseudomonas aeruginosa and hematogenous pulmonary infection with Staphylococcu aureus. Since all models reflect the human clinical conditions, we have achieved good reputation not only in Japan but also in overseas. Using these models, we have investigated the role of mucin in chronic airway infection with P. aeruginosa, mechanisms of action of macrolides drugs against the exerted host and the invading organisms, and pathogenic factors involved in pulmonary infection with S. aureus. Many reports of these studies have been presented at various conferences and published in international journals.

[Chronic respiratory Pseudomonas aeruginosa infection]

1. Kaneko Y., Yanagihara K., Seki M., Kuroki M., Miyazaki Y., Hirakata Y., Mukae H., Tomono K., Kadota J., Kohno S Clarithromycin inhibits overproduction of muc5ac core protein in murine model of diffuse panbronchiolitis Am J Physiol 2003 (in press)

2. Yanagihara K, Tomono K, Kaneko, Miyazaki Y, Tsukamoto K, Hirakata Y, Mukae H, Kadota J-I, Murata I, Kohno S. The role of elastase in a mouse model of chronic respiratory P. aeruginosa infection mimicking diffuse panbronchiolitis J. Med. Microbiol. 2003; 52:531-5

3. Yanagihara K., Tomono K., Imamura K., Kaneko Y., Kuroki M., Sawai T., Miyazaki Y., Hirakata Y., Mukae H., Kadota J., Kohno S. Effect of clarinthromycin on chronic respiratory infection caused by Pseudomonas aeruginosa with biofilm formation J. Antimicrob. Chemother. 2002; 49: 867-70

4. Kaneko Y., Yanagihara K., Kuroki M., Ohi H., Kakeya H., Miyazaki Y., Higashiyama Y., Hirakata Y., Tomono K., Kadota J., Kohno S. Effect of parenterally administered ciplofloxacin in a murine model of pulmonary Pseudomonas aeruginosa infection mimicking ventilator-associated pneumonia. Chemotherapy 2001 Nov-Dec; 47: 421-9.

5. Yanagihara K., Seki M., Cheng P-W. Lipopolysaccharide induces mucus cell metaplasia in mouse lung. Am. J. Respir. Cell Mol. Biol. 2001;24: 66-73

6. Yanagihara K, Tomono K., Kuroki M., Kaneko Y., Sawai T., Ohno H., Higashiyama Y., Miyazaki Y., Maesaki S., Kadota J., Kohno S. Intrapulmonary concentrations of inflammatory cytokines in a mouse model of chronic respiratory infection caused by pseudomonas aeruginosa. Clin Exp Immunol. 2000 122(1): 67-71.

7. Yanagihara K, Tomono K., Sawai T., Kuroki M., Kaneko Y., Ohno H., Higashiyama Y., Miyazaki Y., Hirakata Y., Maesaki S., Kadota J., Tashiro T., Kohno S. Combination therapy for chronic pseudomonas aeruginosa respiratory infection associated with biofilm formation J Antimicrob Chemother. 2000 46: 69-72.

8. Yanagihara K, Tomono K., Sawai T., Hirakata Y., Kadota J., Koga H., Tashiro T., Kohno. Effect of clarithromycin on lymphocytes in chronic respiratory Pseudomonas aeruginosa infection. Am. J Respir Crit Care Med. 1997 155: 337-42.

[Hematogenous pulmonary Staphylococcus aureus infection]

1. Kaneko Y, Yanagihara K, Miyazaki Y, Tsukamoto K, Hirakata Y, Tomono K, Kadota J-I, Tashiro T, Murata I, Kohno S. Efficacy of DQ-113 against methicillin-resistant Staphylococcus aureus and vancomycin-insensitive S. aureus in a model of hematogenous pulmonary infection. Antimicrob. Agents Chemother. 2003 (in press).

2. Yanagihara K, Kaneko Y, Sawai T, Miyazaki Y, Tsukamoto K, Hirakata Y, Tomono K, Kadota J-I, Tashiro T, Murata I, Kohno S. Efficacy of linezolid against methicillin-resistant Staphylococcus aureus and vancomycin-insensitive S. aureus in a model of hematogenous pulmonary infection Antimicrob. Agents Chemother. 2002; 46 (10) 3288-3291.

3. Sawai T, Tomono K., Yanagihara K., Yamamoto Y., Kaku M., Hirakata Y., Koga H., Tashiro T., Kohno S. Role of coagulase in a murine model of hematogenous pulmonary infection induced by intravenous injection of Staphylococcus aureus enmeshed in agar beads. Infect. Immun. 1997 65(2): 466-71.

[Pulmonary Tuberculosis]

Mycobacterium tuberculosis is microorganism that grows relatively slower than other bacteria. It takes 8 weeks to determine whether a specimen contains them, whereas just a couple of days are required for detecting common bacteria. We have been studying the method for detecting a trace amount of M. tuberculosis contained in sputum by detection of its specific gene. As a result, a diagnosis of tuberculosis can now be made in about one day.

Genetic diagnosis is also applied to detection of drug-resistant M. tuberculosis. Generally, drug susceptibility is accessed by the extent of growth inhibition of the organisms by reading the conductivity of the media containing a drug and organisms. Since it takes eight weeks only to detect the M. tuberculosis, the total of 12 weeks is required for susceptibility testing. Rapid determination of drug susceptibility is needed in order to choose the proper drugs at the time of detection of M. tuberculosis. Since it has been revealed that the genetic alterations occurred in particular regions of the specific genes are involved to drug resistance, the molecular technique for analyzing DNA sequence of their specific regions are developed. These advanced techniques are now in available in our section.

1) Fukuda M, Koga H, Ohno H, Yang B, Hirakata Y, Maesaki S, Tomono K, Tashiro T, Kohno S.Relationship between genetic alteration of the rpsL gene and streptomycin susceptibility of Mycobacterium tuberculosis in Japan. J Antimicrob Chemother 1999 43:281-284

2) Yang B, Koga H, Ohno H, Ogawa K, Fukuda M, Hirakata Y, Maesaki S, Tomono K, Tashiro T, Kohno S. Relationship between antimycobacterial activities of rifampicin, rifabutin and
KRM-1648 and rpoBmutations of Mycobacterium tuberculosis. J Antimicrob Chemother. 1998 42(5):621-8.

3) Yang B, Koga H, Ohno H, Ogawa K, Hossain MA, Fukuda M, Hirakata Y, Tomono K, Tashiro T, Kohno S. Detection of Mycobacterium tuberculosis in preserved tuberculous lymph nodes by polymerase chain reaction. Tohoku J Exp Med. 1998 84(2):123-31.

4) Koga H, Miyamoto J, Ohno H, Ogawa K, Tomono K, Tashiro T, Kohno S. A rapid drug susceptibility test for Mycobacterium tuberculosis using the hybridization protection assay. J Antimicrob Chemother. 1997 40(2):189-94.

5) Ohno H, Koga H, Kuroita T, Tomono K, Ogawa K, Yanagihara K, Yamamoto Y, Miyamoto J, Tashiro T, Kohno S. Rapid prediction of rifampin susceptibility of Mycobacterium
tuberculosis. Am J Respir Crit Care Med. 1997 155(6):2057-63.

6) Ohno H, Koga H, Kohno S, Tashiro T, Hara K. Relationship between rifampin MICs for and rpoB mutations of Mycobacterium tuberculosis strains isolated in Japan.
Antimicrob Agents Chemother 1996 Apr;40(4):1053-6

[Research on Fungal Infections]

The research activities of the fungal study group are focused on analysis of pathogenic immunological mechanism of fungal disease, evaluation of new antimycotic drugs, and genetic analysis of mechanisms of resistance to antifungal agents. In clinical practice, since we have treated many patients with fungal disease and all records and information allows us to assess its clinical features and so on. The reputations of both basic and clinical researches have been reported as follows.

CANDIDA

Takashige Miyazaki, et al. Plasma(1-3)-Beta-D-glucan an / antigenemia in patients with candidemia, aspergillosis, and cryptococcosis. J. Clin. Microb. 33: 3115-3118
Kotaro Mitsutake, et al. Enolase Antigen, Mannan Antigen, Cand-Tec Antigen, and b-Glucan in Patients with Candidemia. J. Clin. Microb. 1918-1921

CRYPTOCOCCUS

Yoshihiro Yamamoto, et al. Random Amplified Polymorphic DNA Analysis of Clinically and Enviromentally Isolated Cryptococcus neoformans in Nagasaki. 1995. J. Clin. Microb. 3328-3332
Shigeru Kohno, et al. Epidemiology Studies of Clinical Isolates of Cryptococcus neoformans of Japan by Restriction Fragment Lrngth Polymorphism. 1994. Jpn. Assoc. Infect. Dis. 38: 1512-1517.
Ken-ichi Tanaka, et al. Detection of Cryptococcus neoformans Gene in Patients with Pulmonary Cryptococcosis. 1996. J. Clin. Microb. 34: 2826-2828.

ASPERGILLUS

Shigeru Kohno, et al. A New Antifungal Drug Delivery System, Lipid Nano- Sphere Encasulating Amphotericin B(LNS-AmB), Its Evaluation in The Rat Model of Invasive Pulmonary Aspergillosis. 35Th Interscience Conference on Antimicrobial Agents and Chemotheraphy, 1995. San Francisco, CA
Shigehumi Maesaki, et al. Antifungal activity against Aspergillus fumigatus mycelial cells measured by wet weight, protein concentration and enzymatic activit. J. Mycol. Med. 4: 29-33
H. Yamada, et al. Topical treatment of pulmonary aspergilloma by antifugals. Relationship between duration of the disease and efficacy of therapy. Chest. 103: 1421-1425

IMMUNOLOGY of CRYPTOCOCCUS

Kakeya H, Udono H, Maesaki S, Sasaki E, Kawamura S, Hossain MA, Yamamoto Y, Sawai T, Fukuda M, Mitsutake K, Miyazaki Y, Tomono K, Tashiro T, Nakayama E, Kohno S. Heat
shock protein 70 (hsp70) as a major target of the antibody response in patients with pulmonary cryptococcosis. Clin Exp Immunol. 1999 115(3):485-90.
Sasaki E, Tashiro T, Kuroki M, Seki M, Miyazaki Y, Maesaki S, Tomono K, Kadota J, Kohno S. Effects of macrophage colony-stimulating factor (M-CSF) on anti-fungal activity of
mononuclear phagocytes against Trichosporon asahii. Clin Exp Immunol. 2000 119(2):293-8.

ANALYSIS of ANTI-FUNGAL DRUGS USIBG ANIMAL MODEL

Hossain MA, Maesaki S, Mitsutake K, Kakeya H, Sasaki E, Tomono K, Tashiro T, Kohno S. In-vitro and in-vivo activities of SCH56592 against Cryptococcus neoformans. J Antimicrob Chemother. 1999 44(6):827-9.

Hossain MA, Maesaki S, Razzaque MS, Tomono K, Taguchi T, Kohno S. Attenuation of nephrotoxicity by a novel lipid nanosphere (NS-718) incorporating amphotericin B. J Antimicrob Chemother. 2000 46(2):263-8.

Otsubo T, Maesaki S, Hossain MA, Yamamoto Y, Tomono K, Tashiro T, Seki J, Tomii Y, Sonoke S, Kohno S. In vitro and in vivo activities of NS-718, a new lipid nanosphere incorporating amphotericin B, against Aspergillus fumigatus. Antimicrob Agents Chemother. 1999 43(3):471-5.

Maesaki S, Hossain MA, Miyazaki Y, Tomono K, Tashiro T, Kohno S.

Efficacy of FK463, a (1,3)-beta-D-glucan synthase inhibitor, in disseminated azole-resistant candida albicans infection in mice. Antimicrob Agents Chemother. 2000 44(6):1728-30.

DRUG RESSITANCE in CANDIDA

Maesaki S, Marichal P, Vanden Bossche H, Sanglard D, Kohno S. Rhodamine 6G efflux for the detection of CDR1-overexpressing azole-resistant Candida albicans strains. J Antimicrob
Chemother. 1999 44(1):27-31.

Maesaki S, Marichal P, Hossain MA, Sanglard D, Vanden Bossche H, Kohno S. Synergic effects of tactolimus and azole antifungal agents against azole-resistant Candida albicans strains. J
Antimicrob Chemother. 1998 42(6):747-53.

Miyazaki Y, Geber A, Miyazaki H, Falconer D, Parkinson T, Hitchcock C, Grimberg B, Nyswaner K, Bennett JE. Cloning, sequencing, expression and allelic sequence diversity of ERG3 (C-5 sterol desaturasegene) in Candida albicans. Gene. 1999 5;236(1):43-51.

Miyazaki H, Miyazaki Y, Geber A, Parkinson T, Hitchcock C, Falconer DJ, Ward DJ, Marsden K, Bennett JE. Fluconazole resistance associated with drug efflux and increased transcription of a drug transporter gene, PDH1, in Candida glabrata. Antimicrob Agents Chemother. 1998 42(7):1695-701.

Kakeya H, Miyazaki Y, Miyazaki H, Nyswaner K, Grimberg B, Bennett JE. Genetic analysis of azole resistance in the Darlington strain of Candida albicans. Antimicrob Agents Chemother.
Izumikawa K, Kakeya H, Tsai HF, Grimberg B, Bennett JE. Function of Candida glabrata ABC transporter gene, PDH1. Yeast. 2000 44(11):2985-90.

CLINICAL CASES of FUNGAL DISEASES

Kawamura S, Maesaki S, Tomono K, Tashiro T, Kohno S. Clinical evaluation of 61 patients with pulmonary aspergilloma. Intern Med. 2000 39(3):209-12.

Seki M, Maesaki S, Hashiguchi K, Tomiyama Y, Tomono K, Tashiro T, Kohno S. Aspergillus fumigatus isolated from blood samples of a patient with pulmonary aspergilloma after embolization. Intern Med. 2000 39(2):188-90.

Maesaki S, Kawamura S, Hashiguchi K, Hossain MA, Sasaki E, Miyazaki Y, Tomono K, Tashiro T, Kohno S. Evaluation of sandwich ELISA galactomannan test in samples of positive LA test and positive aspergillus antibody. Intern Med. 1999 38(12):948-50.

Kawamura S, Maesaki S, Omagari K, Hashiguchi K, Tomono K, Tashiro T, Kohno S. Invasive pulmonary aspergillosis diagnosed early by polymerase chain reaction assay. Intern Med. 1999
38(9):744-6.

Kawamura S, Maesaki S, Noda T, Hirakata Y, Tomono K, Tashiro T, Kohno S. Comparison between PCR and detection of antigen in sera for diagnosis of pulmonary aspergillosis. J Clin Microbiol. 1999 37(1):218-20.

CRYPTOCOCCUS GENOME PLAN

http://baggage.stanford.edu/group/C.neoformans/index.html