HBs항원 무반응 SJL/J 마우스에서 F₁(SJL/J*BALB/C) 마우스의 항원전달세포 접종에 의한 항체생성유도

Abstract

[한글]

SJL/J 마우스는 HBs항원/P25에 대한 항체를 생성하지 못하지만, BALB/C와 F^^1 (SJL/J×BALB/C) 마우스는 높은 항체가의 항HBs항체를 생성할 수 있다. 항HBs항체 생성능력이 없는 SJL/J 마우스에는 HBs항원에 특이한, 항원전달세포의 기능상 결손이 존재한다는 시

험관내 실험결과가 보고되어 있다. F^^1 마우스의 항원전달세포는 HBs항원 전달 기능면에서 결손이 없고,SJL/J와 F^^1 (SJL/J×BALB/C) 마우스는 유전적으로 하나의 주요 조직적합 복합체 일배체형이 동일함으로 F^^1 마우스의 항원전달세포와 SJL/J 마우스의 T 세포

간에 항원전달이 가능할 것으로 본다. 따라서 본 연구에서는 F^^1 마우스의 항원전달세포를 시험관내에서 HBs항원으로 처리한후 SJL/J 마우스에 접종함으로써, SJL/J 마우스에서 항HBs항체 생성을 유도하고자 실험을 시행하여 다음의 결과를 얻었다.

1. HBs항원을 복강 또는 정맥내로 접종한 후 효소 면역 측정법으로 혈청내 항HBs항체를 측정한 결과, BALB/C와 F^^1 마우스에서는 항체를 생성하였으나, SJL/J 마우스에서는 항체가 검출되지 않았다. 또한 HBs항원을 족저부에 접종하여 HBs항원 특이 T 림프구의 증식반응 및 IL-2 생성을 측정한 결과, BALB/C 및 F^^1 마우스에서는 증시반응 및 IL-2 생성을 관찰할 수 있었으나, SJL/J 마우스에서는 증식반응이나 IL-2 생성이 관찰되지 않았다.

즉, 시험관내 HBs항원 특이 T 림프구의 증식반응이나 IL-2 생성 여부는 생체내 HBs항체 생성 여부를 반영하였다.

2. 항HBs항체 생성능이 없는 SJL/J 마우스에서 항체 생성을 유도하고자, F^^1 마우스의 항원전달세포를 시험관내에서 HBs항원과 mitomycin C로 전처리하여 SJL/J 마우스에 정맥내로 일차 및 이차접종한 바, 각기 1 :2**5 ∼2**8 및 1 ;2**12 ∼2**15 의 항체가를 관찰하였다. 그러나, BALB/C 마우스의 항원전달세포를 같은 전처리 후 SJL/J 마우스에 접종한 경우에는 항체생성을 관찰할 수 없었다. 또한 항원 처리하지 않은 BALB/C나 F^^1 마우스의 항원전달세포와 HBs항원을 단순 혼합하여 SJL/J 마우스에 정맥 및 복강내 접종한 경우에도 항체생성을 관찰할 수 없었다.

3. 시험관내에서 HBs항원 처리한 F^^1 마우스의 항원전달세포를 SJL/J 마우스에 정맥내로 일차접종 후 10일과 24일에는 항체가가 1 :2**5 ∼2**8 로 검출되었으나, 이차접종을 하지 않으면 일차접종 후 66일에는 항HBs항체를 검출할 수 없었다. 한편, 일차접종 45일

후에 이차접종을 시행할 때 F^^1 마우스의 항원전달세포 없이 HBs항원을 Complete Freund's Adjuvant에 혼합하여 복강내로 접종한 경우에는, 이차접종 후 21일(일차접종 후 66일)에 1 :2**12 ∼2**14 의 항체가를 관찰할 수 있었다.

4. SJL/J 마우스에, 시험관내에서 HBs항원 처리한 후 가열 처리한 항원전달세포를 접종한 다음 HBs항원 특이 T 림프구의 IL-2 생성능을 관찰한 결과, F^^1 마우스의 항원전달세포를 접종한 SJL/J 마우스에서는 IL-2 생성을 관찰할 수 있었으나, SJL/J 및 BALB/C 마우

스의 항원전달세포 투여군과 대조군에서는 IL-2 생성을 관찰할 수 없었다.

이상의 성적으로 보아, HBs항원에 특이한, 항원전달세포의 기능상 결손으로 항HBs항체를 생성하지 못하는 SJL/J 마우스에, SJL/J 마우스와 유전적으로 하나의 동일한 주요조직적합 복합체 일배체형을 가지고 있으며 항HBs항체 생성능이 높은 F^^1 (SJL/J×BALB/C)

마우스의 항원전달세포를 HBs항원으로 시험관내에서 전처리하여 접종함으로써 SJL/J 마우스에서 항HBs항체 생성을 유도할 수 있었다.

Circumvention of nonresponsiveness to HBs antigen in sJL/J mice by the inoculation

of in vitro HBs antigen pulsed antigen presenting cells of F^^1 (SJL/J×BALB/C)

mice

Choong-San Oh

Department of Medical Science The Graduate School, Yonsei University

(Directed by Professor Joo Deuk Kim)

The murine humoral immune response to HBs antigen(Ag) has previously been

demonstrated to be regulated by H-2-linked Ir genes. For example, SJL/J mouse

(H-2**s) does not produce anti-HBs antibody(Ab) after the immunization with

HBSAg/P25, where as BALB/C (H-2**d) and F^^1(SJL/J×BALB/C) mice make high titers

of anti-HBsAb. In vitro experimental data have been accumulated to indicate the

strong possibility that the antigen presenting cells (APC) of SJL/J mice have HBsAg

specific defect(s).

SJL/J and F^^1 mice are genetically haploidentical so that the APC of either mice

can present antigens to the T cells of the other. In addition to this, the APC of

F^^1 mouse are reported to be able to present HBsAg to T cells.

In this communication, we tried to circumvent the HBsAg specific defect in the

APC of SJL/J mouse by immunizing the SJL/J mouse with the F^^1 APC Pulsed in vitro

with HBsAg, and following results were obtained.

1. No anti-HBsAb production, HBsAg specific T cell proliferative response, or the

IL-2 production of HBsAg specific T cells was noted after inoculating SJL/J mice

with HBsAg/P25. On the contrary, in case of BALB/C and F^^1 mice, high titers of

anti-HBsAb production, high HBsAg specific T cell Proliferative responses, and IL-2

production were observed.

2. When the APC of F^^1 mouse after the in vitro HBsAg pulse were injected into

SJL/J mice iv, SJL/J mice Produced as much anti-HBsAb as BALB/C or F^^1 mice did

after the HBsAg injection. But the trials with the BALB/C APC via iv or ip route,

the F^^1 APC via ip route, and the simple mixture of the BALB/C(or F^^1) APC and

HBsAg failed to induce anti-HBsAb in SJL/J mice. And, after the secondary

immunization with HBsAg following the primary one with the HBsAg pulsed F^^1 APC to

SJL/J mice, the same levels of antibodies were produced in SJL/J mice as after the

primary and secondary immunization with the HBsAg pulsed F^^1 APC. Without any

secondary immunization, no antibodies were detected thereafter.

3. The IL-2 production of HBsAg specific T cells was measured after injecting the

HBsAg pulsed APC into footpads of SJL/J mice. The APC were heat treated (45。C for

1 hour) to reduce the allostimulation, if they were from BALB/C or F^^1 mice. Or,

they were treated with mitomycin C, when they were from SJL/J mice. T cells from

SJL/J mice (injected with the HBsAg pulsed & heat treated F^^1 APC) produced IL-2.

But, T cells from all the other SJL/J mice (injected with the HBsAg pulsed &

mitomycin C <or heat> treated SJL/J <or BALB/C> APC) failed to produce IL-2.

These results indicate that the HBsAg specific immunological defect in the APC of

SJL/J mice can be circumvented by immunizing the SJL/J mouse with the F^^1 APC

pulsed with HBsAg in vitro, which are functionally competent and can present HBsAg

to the T cells of SJL/J mice because these two cells share one H-2 haplotype.

Further studies are required to clarify the immunological defects in the hepatitis

B vaccine nonresponders and to apply these techniques to human.

[영문]

The murine humoral immune response to HBs antigen(Ag) has previously been demonstrated to be regulated by H-2-linked Ir genes. For example, SJL/J mouse (H-2**s) does not produce anti-HBs antibody(Ab) after the immunization with HBSAg/P25, where as BALB/C (H-2**d) and F^^1(SJL/J×BALB/C) mice make high titers

of anti-HBsAb. In vitro experimental data have been accumulated to indicate the strong possibility that the antigen presenting cells (APC) of SJL/J mice have HBsAg specific defect(s).

SJL/J and F^^1 mice are genetically haploidentical so that the APC of either mice can present antigens to the T cells of the other. In addition to this, the APC of F^^1 mouse are reported to be able to present HBsAg to T cells.

In this communication, we tried to circumvent the HBsAg specific defect in the APC of SJL/J mouse by immunizing the SJL/J mouse with the F^^1 APC Pulsed in vitro with HBsAg, and following results were obtained.

1. No anti-HBsAb production, HBsAg specific T cell proliferative response, or the IL-2 production of HBsAg specific T cells was noted after inoculating SJL/J mice with HBsAg/P25. On the contrary, in case of BALB/C and F^^1 mice, high titers of

anti-HBsAb production, high HBsAg specific T cell Proliferative responses, and IL-2 production were observed.

2. When the APC of F^^1 mouse after the in vitro HBsAg pulse were injected into SJL/J mice iv, SJL/J mice Produced as much anti-HBsAb as BALB/C or F^^1 mice did after the HBsAg injection. But the trials with the BALB/C APC via iv or ip route, the F^^1 APC via ip route, and the simple mixture of the BALB/C(or F^^1) APC and HBsAg failed to induce anti-HBsAb in SJL/J mice. And, after the secondary immunization with HBsAg following the primary one with the HBsAg pulsed F^^1 APC to SJL/J mice, the same levels of antibodies were produced in SJL/J mice as after the primary and secondary immunization with the HBsAg pulsed F^^1 APC. Without any secondary immunization, no antibodies were detected thereafter.

3. The IL-2 production of HBsAg specific T cells was measured after injecting the HBsAg pulsed APC into footpads of SJL/J mice. The APC were heat treated (45。C for 1 hour) to reduce the allostimulation, if they were from BALB/C or F^^1 mice. Or,

they were treated with mitomycin C, when they were from SJL/J mice. T cells from SJL/J mice (injected with the HBsAg pulsed & heat treated F^^1 APC) produced IL-2.

But, T cells from all the other SJL/J mice (injected with the HBsAg pulsed & mitomycin C <or heat> treated SJL/J <or BALB/C> APC) failed to produce IL-2.

These results indicate that the HBsAg specific immunological defect in the APC of SJL/J mice can be circumvented by immunizing the SJL/J mouse with the F^^1 APC pulsed with HBsAg in vitro, which are functionally competent and can present HBsAg to the T cells of SJL/J mice because these two cells share one H-2 haplotype.

Further studies are required to clarify the immunological defects in the hepatitis B vaccine nonresponders and to apply these techniques to human.