The paper deals with the process of acquiring professional vocabulary and grammar in English for Specific Purposes (ESP) in pre-service teacher preparation. The main objective of the research was to (1) discover how much students learn in the ICT-enhanced process of instruction, where autonomous work and collaborative learning are applied so that to build preconcepts and re-construct misconceptions in ESP; (2) detect the sources students exploited towards acquiring the learning content. The research was conducted at the Faculty of Education, University of Jan Evangelista Purkyne, Usti nad Labem, Czech Republic. Research sample consists of 62 probands enrolled in bachelor and master study programmes English Language and Literature and Teaching English Language and Literature. The research process is structured into three phases, and didactic tests are applied after each of them. Moreover, types of sources students used for acquiring the learning content were monitored and analyzed. Six hypotheses were tested and results showed statistically significant increase in knowledge after each phase.