Background: In Ethiopia, first line antiretroviral therapy failure is growing rapidly. However, unlike first line therapy, to date, very little is known about the outcomes of second line therapy. Thus, this study assessed the rate of viral re-suppression and retention to care and their predictors among HIV infected patients on second line therapy. Methods: A retrospective cohort study was conducted among 642 HIV infected patients on second line therapy at Dessie Comprehensive Specialized Hospital, northeast Ethiopia from October 2016 to November 2019. Proportional Cox regression model was computed to explore predictors of viral re-suppression (Viral load less than 1000 copies/mL) and attrition to care. Results: Out of 642 subjects, 19 (3%), 44(6.9%), 70(10.9%), and 509(79.3%) patients were lost to follow up, died, transferred out, and alive on care, respectively. Similarly, 82.39 % (79.24 - 85.16%) patients had achieved viral re-suppression, with 96 per 100-person year rate of re-suppression in 550.16 year follow up. Patients who switched timely to second line therapy were at a high rate of viral re-suppression than patients who delayed to switch [Adjusted Hazard Rate, AHR = 1.43 (95%CI: 1.17-1.74)]. Not having drug substitution history [AHR =1.25 (95%CI: 1.02-1.52)] was positively associated with viral re-suppression, in contrast, being on anti-TB treatment [AHR = 0.67 (95% CI: 0.49 – 0.91)] negatively associated with viral re-suppression. By excluding transferred out cases, 63 (11%, 95% CI: 9.6 – 13.8%) out of 572 patients were failed to retain on care with 7.1 per 100 person year rate of attrition in 887.25 year observation. Patients who were ambulatory or bedridden at the time of therapy switch were more at risk of attrition to care as compared with workable patients [Adjusted Hazard Rate, AHR= 2.61 (95% CI: 1.40-4.87]. Similarly, being not virally re-suppressed [AHR= 6.87 (95% CI: 3.86-12.23)], CD4 count < = 450 cells/mm3 [AHR= 2.61 (95% CI: 1.40-4.87] were also positively associated with attrition to care. Conclusions: A significant number of patients had failed to achieve viral re-suppression and to retain on care. Most identified factors related to patient monitoring. Hence, patient centered intervention should be strengthen, besides treatment switch.