despite、despite of、in spite of的区别为：意思不同、用法不同、侧重点不同。
一、意思不同
1.despite：即使，尽管，尽管(自己)不愿意。
2.despite of：尽管。
3.in spite of：尽管；不管，不顾。
二、用法不同
1.despite：despite书面用词，指不受某事或某种原因的阻碍，语气轻于in spite of。
2.despite of：in spite of在这些词中语气最强，可与despite换用，口语或书面语中均可使用。
3.in spite of：语气最弱，仅表示一种障碍的存在。
三、侧重点不同
1.despite：侧重于表示的感觉更随意。
2.despite of：不要用据说是古用法现在有考试算错误。
3.in spite of：侧重于表示的感觉更严谨。
We went swimming in spite of rain. (尽管下雨，我们还是去游泳了) (正)
We went swimming in spite rain. (误)
We went swimming inspite of rain. (误)Despite all his efforts Robert failed. (虽然罗伯特尽了一切努力，还是失败了) (正)
Despite of all his efforts Robert failed. (误)
与其他所有介系词 (片语) 一样，in spite of 和 despite 後面通常接名词 (片语)，但亦可接动名词。例如：
Rock managed to eat a substantial lunch in spite of having eaten a big breakfast. (尽管早餐吃了很多，洛克午餐还是大吃一顿)In spite of being very tired, I stayed awake. (尽管非常疲倦，我仍保持清醒)James doesn’t lead a life of luxury despite making a lot of money. (尽管赚了很多钱，詹姆斯的生活并不奢华)Despite being sick, Grace went to work. (尽管生病，葛蕾丝还是去工作)
In spite of 和 despite 亦可接 that 所引导的名词子句当受词。根据文法，介系词後面不能直接接 that 子句当受词，但我们可以在 that 子句的前面加上 the fact，使 the fact 作为介词的受词，这样其後的 that 子句就成了 the fact 的同位语，百分之百合乎语法要求。The fact that 中的 the fact 虽具有举足轻重的语法功能，但本身并无意义。
The fact 与 that 所引导的名词子句为同位语。原本引导名词子句的 that 是连接词，在加上 the fact 当同位语後，that 变成关系代名词。例如：
In spite of the fact that I was very tired, I stayed awake. (尽管非常疲倦，我仍保持清醒)Despite the fact that she was sick, Grace went to work. (尽管生病，葛蕾丝还是去工作)We enjoyed ourselves very much, in spite of the fact that we had no money. (我们虽然没有钱，但仍然过得很愉快)She still loves him, despite the fact that he left her. (尽管他离开了她，她仍然爱他)
In spite of 和 despite 皆可接 oneself 来表示「不由自主地，忍不住地」的意思。例如：
Mary started to scream, almost in spite of herself. (玛丽几乎不由自主地尖叫起来)John laughed despite himself. (约翰忍不住笑了起来)She picked her nose in public, despite herself. (她忍不住当众挖鼻孔)
