장티푸스환자의 중성백혈구의 nitroblue tetrazolium 색소환원능력에 관한 연구

Abstract

[한글]

Nitroblue Tetrazolium Test and Stimulated Test in Typhoid Fever

Kyung-Suk Kim, M.D.

Department of Medical Science The Graduate School, Yonsei University

(Directed by Prof. Eung Suk Chai, M.D.)

Nitroblue tetrazolium (N. B. T.) dye has been widely used in histochemical

studies of various enzymes. At the sits of enzyme activity, colorless tetrazolium

is reduced to a water-insoluble formazan which can easily be recognized as a deep

blue deposit.

Since Holmes et al. (1966) described patients whose neutrophils are capable of

engulfing but unable to kill the bacteria and demonstrated N. B. T. dye could not

be reduced to formazan in these patients, it hart been suggested that there is

close relationship between the ability of neutrophils to reduce N. B. T dye and to

kill the bacteria after phagocytosis.

Although the mechanism of reduction of N. B. T. to form formazan in phagocytic

neutrophilsis unclear, Park et al. (1968) reported that the number of neutophils

reducing N. B. T dye in vitro was significantly elevated in patients with acute

systemic bacterial infection. Since then N. B. T. test has been used as a means of

differentiating bacterial from non-bacterial infection in febrile patients (Park et

al., 1968; Feigin., 1971, Humbert et al., 1971). It was, however, learned later

that N. B. T. values were not increased in the presence of bacterial infection if

corticosteroid was given or complicated by chronic vascular disease, diabetes

mellitus, influenza, and alcoholic intoxication. Local infection and chronic

infection are also known to be showing normal N. B. T. values though there is

considerable controversy on this issue(Humbert et al., 1971).

In 1973, Kim observed high N.S. T. values in patients with typhoid fever and

postulated that typhoid bacilli and/or their products might play significant role

in activating neutrophils as well as mononuclear cells in reticuloendothelial

system. On the contrary, Lim et al. (1973) reported N.B.T positive neutrophils are

net increased in patients with typhoid fever through the same experiment. There has

been no explanation for these different observaton yet. Grush and Mauer(1969)

showed instant rapid rise of N. B. T. positive neutrophils when typhoid vaccines

were given to healthy people.

Park (7971) also introduced a simple technique testing intrinsic defect of

neutrophils to phagocytize and kill bacteria, when N. B. T. values are not

increased in the presence of acute bacterial infection, by observing significant

rise in N. B. T positive neutrophils when Escherichia coli endotoxin is added in

vitro.

The purpose of the present study was to determine if N. B. T. positive

neutrophils are increased in typhoid fever as in other bacterial infection and

there is intrinsic defect of neutrophils in phagocrtizing and killing bacteria, by

performing stimulates N. B. T test.

N. B. T, and stimulated N. B. T. test with Escherichia coli and Salmonella typhi

endotoxin was performed on 27 healthy adults and 35 patients with typhoid fever by

the same method of Park and Park (1968, 1971). The summary of the results are as

following.

1. There was no significant difference in white blood cell count and absolute

number of neutrophils between normal control and typhoid fever, mean value toeing

6.660±331/㎜, 3,851±262/㎣ and 5,280±334/㎣, 3,384±323/㎣ respectively.

2. Though the mean percentage of N. B. T. positive neutrophils in typhoid fever

was slightly higher than control(typhoid fever: 5.5±0.6, control: 3.4±0.51, there

was no statistically significant difference(P>0.05).

3. Significant increase in number of N. B. T positive neutrophils was observed

when it was stimulated by Escherichia coli and Salmonella typhi endotoxin in vitro

(control with E. coli endotoxin: 13.2±1.5%, control with Salmonella typhi

endotoxin: 13.7±1.9%, typhoid fever with E. coli endotoxin: 21.6±2.1% and typhoid

fever with Salmonella typhi endotoxin: 20.1±1.7%).

4. There was no meaningful difference in the degree of stimulation, either

between control and typhoid fever or between E. soli endotoxin and Salmonella typhi

endotoxin.

It might be concluded that while the number of neutrophils reducing N. B. T. dye

in typhoid fever is not elevated as in other bacterial infection, there appears to

be no intrinsic defect in neutrophils to phagocytize and kill bacteria. The

possible mechanism of failure to increase N. B. T_values in typhoid fever was

discussed.

[영문]

Nitroblue tetrazolium (N. B. T.) dye has been widely used in histochemical studies of various enzymes. At the sits of enzyme activity, colorless tetrazolium is reduced to a water-insoluble formazan which can easily be recognized as a deep blue deposit.

Since Holmes et al. (1966) described patients whose neutrophils are capable of engulfing but unable to kill the bacteria and demonstrated N. B. T. dye could not be reduced to formazan in these patients, it hart been suggested that there is close relationship between the ability of neutrophils to reduce N. B. T dye and to kill the bacteria after phagocytosis.

Although the mechanism of reduction of N. B. T. to form formazan in phagocytic neutrophilsis unclear, Park et al. (1968) reported that the number of neutophils reducing N. B. T dye in vitro was significantly elevated in patients with acute systemic bacterial infection. Since then N. B. T. test has been used as a means of differentiating bacterial from non-bacterial infection in febrile patients (Park et al., 1968; Feigin., 1971, Humbert et al., 1971). It was, however, learned later that N. B. T. values were not increased in the presence of bacterial infection if

corticosteroid was given or complicated by chronic vascular disease, diabetes mellitus, influenza, and alcoholic intoxication. Local infection and chronic infection are also known to be showing normal N. B. T. values though there is

considerable controversy on this issue(Humbert et al., 1971).

In 1973, Kim observed high N.S. T. values in patients with typhoid fever and postulated that typhoid bacilli and/or their products might play significant role in activating neutrophils as well as mononuclear cells in reticuloendothelial system. On the contrary, Lim et al. (1973) reported N.B.T positive neutrophils are net increased in patients with typhoid fever through the same experiment. There has been no explanation for these different observaton yet. Grush and Mauer(1969) showed instant rapid rise of N. B. T. positive neutrophils when typhoid vaccines were given to healthy people.

Park (7971) also introduced a simple technique testing intrinsic defect of neutrophils to phagocytize and kill bacteria, when N. B. T. values are not increased in the presence of acute bacterial infection, by observing significant rise in N. B. T positive neutrophils when Escherichia coli endotoxin is added in

vitro.

The purpose of the present study was to determine if N. B. T. positive neutrophils are increased in typhoid fever as in other bacterial infection and there is intrinsic defect of neutrophils in phagocrtizing and killing bacteria, by performing stimulates N. B. T test.

N. B. T, and stimulated N. B. T. test with Escherichia coli and Salmonella typhi endotoxin was performed on 27 healthy adults and 35 patients with typhoid fever by the same method of Park and Park (1968, 1971). The summary of the results are as following.

1. There was no significant difference in white blood cell count and absolute number of neutrophils between normal control and typhoid fever, mean value toeing 6.660±331/㎜, 3,851±262/㎣ and 5,280±334/㎣, 3,384±323/㎣ respectively.

2. Though the mean percentage of N. B. T. positive neutrophils in typhoid fever was slightly higher than control(typhoid fever: 5.5±0.6, control: 3.4±0.51, there was no statistically significant difference(P>0.05).

3. Significant increase in number of N. B. T positive neutrophils was observed when it was stimulated by Escherichia coli and Salmonella typhi endotoxin in vitro (control with E. coli endotoxin: 13.2±1.5%, control with Salmonella typhi

endotoxin: 13.7±1.9%, typhoid fever with E. coli endotoxin: 21.6±2.1% and typhoid fever with Salmonella typhi endotoxin: 20.1±1.7%).

4. There was no meaningful difference in the degree of stimulation, either between control and typhoid fever or between E. soli endotoxin and Salmonella typhi endotoxin.

It might be concluded that while the number of neutrophils reducing N. B. T. dye in typhoid fever is not elevated as in other bacterial infection, there appears to be no intrinsic defect in neutrophils to phagocytize and kill bacteria. The

possible mechanism of failure to increase N. B. T_values in typhoid fever was discussed.